СПОСОБ ОПРЕДЕЛЕНИЯ АКТИВНОСТИ ЩЕЛОЧНОЙ ФОСФАТАЗЫ В МАЗКАХ ГЕМОЛИМФЫ У ПЧЕЛ

Изобретение относится к области ветеринарной медицины. Способ заключается во взятии мазков, их обработки, фиксации, заливки и инкубировании в среде, состоящей из буферного раствора, нафтилофосфата и красителя нейтрального прочного синего В, дополнительном окрашивании нейтральной красной краской, оценке качественной окраски. Способ позволяет уменьшить материальные затраты и время. 5 ил., 1 табл.

ПЛАЗМИДНЫЙ ВЕКТОР И СПОСОБ ВЫЯВЛЕНИЯ НОНСЕНС-МУТАЦИЙ И МУТАЦИЙ СДВИГА РАМКИ СЧИТЫВАНИЯ В ГЕНЕ BRCA1

Настоящее изобретение относится к области молекулярной биологии. Предложен способ выявления в гене BRCA1 мутаций сдвига рамки считывания и нонсенс-мутаций, заключающийся в создании рекомбинантных плазмид, в которых амплифицированный фрагмент гена находится в единой трансляционной рамке с геном щелочной фосфатазы Е.соli (phoA). Сконструирован плазмидный вектор pPhoA-frame, который содержит последовательность ДНК, кодирующую щелочную фосфатазу E.coli. Внутрь этой последовательности встроен фрагмент ДНК, содержащий сайты узнавания эндонуклеаз рестрикции BglII, StuI, ApaI, SacII и предназначенный для клонирования фрагментов гена BRCA1 (полилинкер). Амплифицированный фрагмент гена BRCA1 встраивается в плазмидный вектор pPhoA-frame в единой трансляционной рамке с phoA. Наличие в исследуемом фрагменте гена мутаций, нарушающих целостность рамки считывания, оценивается визуально по отсутствию окраски колоний клеток E.coli, трансформированных полученной рекомбинантной плазмидой, на индикаторной чашке ...

СПОСОБ ОПРЕДЕЛЕНИЯ НАЛИЧИЯ СИБИРЕЯЗВЕННОЙ ИНФЕКЦИИ В БИОЛОГИЧЕСКИХ ЖИДКОСТЯХ И ОКРУЖАЮЩЕЙ СРЕДЕ

Способ детекции сибиреязвенного патогена в окружающей среде и биологических жидкостях основан на функциональной детекции активности одного из компонентов сибиреязвенного токсина, слабой протеазы летального фактора сибирской язвы (LF). Примененный принцип высокочувствительной детекции протеолитической активности LF базируется на системе амплификации сигнала, происходящего от акта разрезания субстрата, при помощи находящегося в составе субстрата вспомогательного фермента щелочной фосфатазы Escherichia coli (АР). В составе рекомбинантного субстрата LF помимо щелочной фосфатазы и специфической для LF субстратной последовательности RRKKVYPYPME находятся пептид, биотинилирующийся in vivo и in vitro при помощи биотин-лигазы E.coli и обеспечивающий иммобилизацию субстрата на твердой фазе (поверхности плашек) за счет взаимодействия с авидином и его производными, и последовательности шести гистидинов для очистки рекомбинантного субстрата из периплазмы E.coli при помощи металл-хелатной смолы. Способ ...

СПОСОБ ОПРЕДЕЛЕНИЯ ЭЛЕКТРОНОНАСЫЩЕННОГО АРОМАТИЧЕСКОГО АМИНА, ЯВЛЯЮЩЕГОСЯ МАРКЕРОМ СОДЕРЖАНИЯ АНАЛИТА В БИОЛОГИЧЕСКОЙ ЖИДКОСТИ, И СРЕДСТВО ДЛЯ ЕГО ОСУЩЕСТВЛЕНИЯ

Использование: в медицине, для определения аналита при помощи образования гетерополисинего. Сущность: аналит превращают с веществом, которое приводит к электрононасыщенному амину, который вступает в контакт с труднорастворимой солью гетерополикислоты. Средство для осуществления способа содержит труднорастворимую соль гетерополикислоты в количестве, достаточном для перевода всего амина в гетерополисиний. 2 с и 2 з. п. ф-лы, 7 ил., 4 табл.

КОМПОЗИЦИИ ДЛЯ РЕАКЦИИ ОБРАТНОЙ ТРАНСКРИПЦИИ С ГОРЯЧИМ СТАРТОМ ИЛИ ДЛЯ ПОЛИМЕРАЗНОЙ ЦЕПНОЙ РЕАКЦИИ С ОБРАТНОЙ ТРАНСКРИПЦИЕЙ С ГОРЯЧИМ СТАРТОМ

... 1. Композиция для реакции обратной транскрипции с горячим стартом, которая содержит ион Mg, четыре типа dNTP, обратную транскриптазу, пирофосфат (PPi), и фосфатазу (ППазу).2. Композиция для реакции обратной транскрипции с горячим стартом по п. 1, в котором композиция, содержит пирофосфат (PPi) в концентрации 0,1-5 мМ, а пирофосфотазу в количестве 0,005-0,25 Ед.3. Композиция для реакции обратной транскрипции с горячим стартом по п. 1, дополнительно содержащая один или несколько праймеров для обратной транскрипции.4. Композиция для реакции обратной транскрипции с горячим стартом по п. 1, где композиция заморожена или высушена.5. Композиция для реакции обратной транскрипции с горячим стартом по п. 1, дополнительно содержащая нуклеотидную матрицу.6. Композиция для реакции обратной транскрипции с горячим стартом по п. 5, в котором нуклеотидная матрица является РНК.7. Композиция для реакции обратной транскрипции с горячим стартом по п. 1, дополнительно содержащая краситель, который не реагирует ...

СПОСОБ ОПРЕДЕЛЕНИЯ АКТИВНОСТИ ЩЕЛОЧНОЙ ФОСФАТАЗЫ В МАЗКАХ ГЕМОЛИМФЫ У ПЧЕЛ

Способ определения активности щелочной фосфатазы в мазках гемолимфы у пчел, включающий подготовку мазков биологического субстрата, взятие обработки, фиксацию, промывку, заливку и их инкубирование в среде, состоящей из буферного раствора дистиллированной воды, нафтилофосфата и нейтрального прочного синего В, промывание, дополнительное окрашивание прочным красным, оценку качественной окраски на щелочную фосфотазу по окраске, отличающийся тем, что в качестве биологического субстрата используют мазки гемолимфы пчел, а для приготовления буферной смеси используют 0,04 М раствор 5,5-диэтилбарбитурат натрия (веронал натрия) - С8Н11N2О3 Na и 0,1 М раствор соляной кислоты (HCl) 1,13-1,65 мл, для обеспечения рН 9,0-9,2.

Способ приготовления инкубационной среды для потенциометрического определения активности @ -АТФ-азы

... 1. СПОСОБ ПРИГОТОВЛЕНИЯ ИНКУБАЦИОННОЙ СРЕДЫ Д)1Я ПОТЕНЦИОМЕТРИЧЕСКОГО ОПРЕДЕЛЕНИЯ АКТИВНОСТИ Н -АТФ-АЗЫ, включающий взвешивание компонентов, растворение их в дистиллированной воде,, доведение рН раствора до 8,3iO,1, отличающийся тем, что, с целью повышения точности определения активности фермента, для растворения компонентов используют дистиллированную воду, предварительно обработанную постоянным электрическим током в электролизере с полупроницаемой мембраной при напряжении на электродах 300-600 В и силе тока 10-30 мА в течение 5-30 мин, при этом использую.т воду из катодной полови- . ны электролизера, а доведение рН среды осуществляют добавлением воды i из обеих половин электролизера, после чего инкубационную среду защищают слоем жидкого индифферентного гидрофобного газонепроницаемого вещества . 2. Способ non.t, отличающийся тем, что для запреты инкубационной Среды используют вазелин. Слд СО СП СО 4 ...

VERWENDUNG EINES SCHWER LOESLICHEN SALZES EINER HETEROPOLYSAEURE ZUR BESTIMMUNG EINES ANALYTS, ENTSPRECHENDES BESTIMMUNGSVERFAHREN SOWIE HIERFUER GEEIGNETES MITTEL

NACHWEIS VON PROSTAT-SPEZIFISCHEN ANTIGEN IN BRUSTTUMOREN

CHEMOLUMINESZENTE 3-(SUBSTITUIERTE ADAMANT-2'-YLIDENE)1,2-DIOXETANE

VERFAHREN ZUR FAERBUNG VON NICHTRADIOAKTIVEN MARKIERTEN NUKLEINSAEUREN UND ZUSAMMENSETZUNG ZUM GEBRAUCH IN DIESEM VERFAHREN.

Chromogene Dibenzoxazepinon- und Dibenzothiazepinon-Enzymsubstrate.

Sonden

VERFAHREN ZUR IDENTIFIZIERUNG EINZELNER AKTIVER EINHEITEN AUS KOMPLEXEN GEMISCHEN

Stabilisierte Enzym-Mischungen.

Stabilisierte Enzym-Mischungen.

Reagenz und Verfahren zur Bestimmung von Kupplungsverbindungen

Homogeneous fluorescent assay for kinase, phosphatase or phosphodiesterase, useful in screening for pharmaceuticals and plant-protection agents, uses polycationic polymer as quencher

Homogeneous assay for quantitative measurement of kinase, phosphatase or phosphodiesterase reactions by reacting the enzyme with a fluorescent, (de)phosphorylatable substrate (A) in presence of polycationic polymer (B) containing quencher groups. The change in phosphorylation is determined from a change in fluorescence.

VERFAHREN UND MITTEL ZUR IMMUNOLOGISCHEN BESTIMMUNG VON ENZYMEN

VERFAHREN ZUM NACHWEIS VON SUBSTANZEN

FLUOROGENE PHOSPHORSAEUREESTER, VERFAHREN ZU DEREN HERSTELLUNG SOWIE VERFAHREN UND MITTEL ZUM NACHWEIS UND ZUR FLUORIMETRISCHEN BESTIMMUNG VON PHOSPHATASEN

FLUORESZENZPOLARIZATIONSBESTIMMUNGEN UNTER VERWENDUNG VON POLYIONEN

Chemiliumineszierendes Substrat von Hydrolasen wie Phosphatasen

TESTVERFAHREN ZUR MESSUNG DER PHOSPHORYLIERUNG

TOXINS AND ANTIBODIES OF C. DIFFICILE

Antibodies (Ab) specific for toxins of Clostridium difficile, opt. bound to a solid support, are new. Esp. Ab are monospecific for either A or B toxin. Also new are assays for toxigenic C. difficile using Ab. The immobilised Ab is incubated with the test sample to bind C. difficile antigens. The bound antigen is then incubated with labelled, monospecific Ab, and the presence of label determined by detecting presence of the label. Particularly, the label used is alkaline phosphatase. In a modification, the monospecific Ab is not labelled, and detection is by reaction with a second, labelled antibody specific for the first. Alternatively, toxins are detected by agglutination of appropriately sensitised solid particles (or by inhibition of such agglutination).

Stabilisierte flüssige Mischungen für die Markierung von Nukleinsäuren

Hybridisation assay

Method of testing

CHROMOGENIC AND FLUOROGENIC ESTERS

DETECTION AND IMAGING IN BIOCHEMICAL ASSAYS USING PHOSPHOR SCREENS

Monitoring bacterial contamination of a wound involving assay of adenosine triphosphate

The invention provides a method of monitoring the bacterial contamination of a wound comprising monitoring the adenosine triphosphate (ATP) concentration of wound fluid removed from the wound. Preferably, an enzyme-coupled reaction such as the luciferin-luciferase reaction is used for the monitoring. Diagnostic kits and wound dressings adapted for use in the method are also provided.

Phosphatese detection using 4-acetamidophenyl phosphate substrate

Phosphatase activity is detected electrochemically with 4-acetamidophenyl phosphate (paracetamol phosphate) as substrate.

Pharmaceutical compositions containing fluorinated or perfluorinated carboxylic acids

REAGENT AND METHOD FOR THE DETERMINATION OF HUMAN-MUSCLE ADOLASE

Coumarin snd quinolinone derivatives

Compounds of the formula wherein Y is M is hydrogen, or an alkali metal or ammonium ion, X is -O- or an amino group, R is H, Cl, Br, CN or carbamoyl, R1 is H or -SO3H, and A is one of several particular organic radicals, are especially useful in the photometric or fluorimetric determination of phosphatase or sulphatase activity in a liquid sample.

Screening Assay

Methods for measuring protein kinase and phosphatase activity

Paracetamol phosphate for immunoassays

Materials and methods relating to the diagnosis and treatment of pre-eclampsia

Inositol phosphoglycans for therapeutic use in the treatment of diabetes and obesity

Conjugates of amino sugars with steroids

Conjugates of amino sugar-steroid hormone are prepared by binding an amino sugar to steroid hormone at 3- or 17-position by a direct reaction or by binding with a binding agent, and the conjugate is useful in an enzyme immunoassay method.

chemiluminescent 3-(substituted adamant-2'-ylidene) 1,2-dioxetanes

Reverse transcriptase assay kit, use thereof and method for analysis or RT activity in biological samples.

Chemical assay technique of the phosphatasic enzymes allowing to determine the diseases with retrovirus: paludism with répitition hepatitis ictère AIDS and primary education cancer of the liver and to differentiate their seropositivity

PROTEIN KINASE NPK-110

SIMPLIFIED SEQUENZIELLE CHEMOLUMINISZENZ MEASUREMENT

METHOD AND COMPOSITIONS TO IMPROVED POLYNUCLEOTIDSYNTHESE

NEW KCNQ POLYPEPTIDE AND THEIR USE WITH THE DIAGNOSIS OF MENTAL DISORDERS

PROCEDURE FOR THE IDENTIFICATION OF AT PHOSPHOINOSITID OBTAINING SIGNALLING CONNECTIONS TAKEN PART AS WELL AS USE OF IT WITH THE PRODUCTION OF DRUGS

PROCEDURE AND KIT FOR THE ANALYSIS OF A GOAL OF NUCLEINSÄUREFRAGMENTES

TEST PROCEDURE FOR THE MEASUREMENT OF THE PHOSPHORYLATION

PROCEDURE AND COMPOSITIONS IN CONNECTION WITH GENE SILENCING

PROCEDURE FOR the PRODUCTION of a HETERO DIMER DERIVATIVE of the ENZYME PROTEINPHOSPHATASE TYPE 2A

VERFAHREN ZUM NACHWEIS VON SUBSTANZEN

PROOF OF IONS IN LIQUIDS

PROCEDURE FOR THE REGULATION OF ALKALINE PHOSPHATASE UNDER REMOVAL OF HEMOGLOBIN DISTURBANCES

PROOF OF DNA BINDUNGSPROTEINEN

FLUORESCENCE SUBSTRATES TO THE PROOF OF ORGANOPHOSPHATASE ENZYMAKTIVITÄT

PROCEDURE FOR THE PROOF OF ACETYLASE OR DEACETYLASE ACTIVITY IN A IMMUNOASSAY

PROCEDURE FOR MEASURING GROUPS CHEMICAL TO BIOLOGICAL MOLECULES BOUND ONES,

DIRECT PROOF OF MOLECULES IN POLYACRYLAMIDGEL

ENZYMATIC ANALYSIS USING A SUBSTRATE THAT NIERDERSCHLAG RESULTS IN A FLUORESCENT ONE

STRAND DISPLACEMENT AMPLIFICATION.

FLUOROKETONE AS BLOCKED ENZYME INHIBITORS FOR USE IN IMMUNOASSAYS

PROCEDURE FOR the REINFORCEMENT of the CHEMILUMINESZENZ OF ENZYME-TRIGGERED 1,2-DIOXETANEN AND ITS USE

PROCEDURE FOR THE DETERMINATION OF THE CILIOSTATI FACTOR WITH CYSTI FIBROSE.

FLUOROMETRI TEST PROCEDURE REAGENTS SUITABLE BY ALLERGI REACTIONS AND FOR IT.

REGULATION METHOD INCLUDING A PHASE SEPARATION AND A TEST RECORD FOR THIS.

DERIVATISIERTE NUCLEINSAEURE SEQUENCE, PROCEDURE FOR THEIR PRODUCTION AS WELL AS THEIR USE FOR THE PROOF OF NUCLEINSAEUREN.

PROCEDURE FOR THE REGULATION OF ISO ENZYMES.

USE OF DERIVATISIERTER ALKALINE PHOSPHATASE AS STANDARD.

1-NAPHTHOLPHTHALEINMONOPHOSPHATE, PROCEDURE FOR THEIR PRODUCTION AS WELL AS THEIR USE FOR the REGULATION the ALKALINE PHOSPHATASE.

REGULATION OF POTASSIUM IONS IN LIQUIDS.

PROCEDURE AND REAGENT FOR THE REGULATION THE CREATINKINASE.

SUBSTRATE BEGINNING OF MIXTURES IN a 2-AMINO2-METHYL-1-PROPANIL CONTAINING BUFFER FUER ALKALIN PHOSPHATASE SAMPLE.

RECOMBINED ALKALINE PHOSPHATASE FROM CALF INTESTINE

PARACETAMOLPHOSPHAT FOR IMMUNOASSAYS

USE OF A WITH DIFFICULTY SOLUBLE SALT HETEROPOLYSÄURE FOR THE DETERMINATION OF A ANALYTS, EQUIVALENT REGULATION PROCEDURE AS WELL AS FOR THIS SUITABLE MEANS.

PRODUCTION AND USE OF FLUORESCENT BENZOTHIAZOLDERIVATEN

USE FROM SUBSTANCES TO PREVENTING OF ENDOTOXINSCHOCK

IMMUNOCHEMI PROCEDURE AND TEST KIT FOR THE REGULATION THE SOUR PROSTATAPHOSPHATASE.

PROCEDURE AND REAGENT FOR THE DIFFERENTIATED REGULATION OF ISO ENZYMES THE ALKALINE PHOSPHATASE.

FUNCTIONAL PROOF OF LIPOPROTEIN WITH HIGH DENSITY

PROCEDURE AND DEVICE FOR THE QUANTITATIVE ONE CHROMATOGRAPHIERUNG.

DIISOPROPYLFLUOROPHOSPHATASE AS WELL AS THEIR USE AND PRODUCTION

CONNECTIONS, COMPOSITIONS AND METHODS TO THE PRODUCTION OF CHEMILUMINESZENZ WITH PHOSPHATASE ONES ENZYMES

PRESERVATION OF DIAGNOSTIC TESTS

PROCEDURE FOR THE PROOF OF INTERFERON ACTIVITY

ANTI- MIKROBI AGENTS AND SCREENING PROCEDURES FOR IT

EUROPIUM AND TERBIUM CHELATORE FOR TIME-SOLVED FLUOROMETRI TEST

ALIZARINE DERIVATIVES AS NEW ONE CHROMOGENE SUBSTRATES

PROCEDURE AND TEST KIT FOR THE DETERMINATION OF THE 5 ' - NUKLEOTIDASE ACTIVITY

NEP INHIBITORS FOR THE TREATMENT OF FEMALE SEXUAL DISTURBANCES

CHEMILUMINESZENZ SUBSTRATES OF HYDROLYTIC ENZYMES SUCH AS PHOSPHATASEN

NEW SUBSTRATES FOR THE DETECTION OF MIKROBIELLEN METABOLITES

DIAGNOSTIC METHOD IS PRODUCED BASED ON THE FUNCTION OF A PROTEIN IN OF A CELLFREE ENVIRONMENT, ON THE BASIS OF A SAMPLE OF NUCLEIC ACIDS

PROCEDURE FOR THE IDENTIFICATION OF INDUCTORS AND RESTRICTORS OF PROGRAMMED CELL DEATH

FLUORESZENZPOLARIZATIONSBESTIMMUNGEN USING POLYIONEN

PROTEIN KINASE NPK-110

PROTEIN KINASE NPK-110

Methods of diagnosing cervical cancer

The invention provides reagents and methods for detecting pathogen infections in human samples. This detection utilizes specific proteins to detect the presence of pathogen proteins or abnormal expression of human proteins resulting from pathogen infections. Specific methods, compositions and kits are disclosed herein for the detection of oncogenic Human papillomavirus E6 proteins in clinical samples.

MASS SPECTROMETRY METHOD FOR MEASURING THIAMINE IN BODY FLUID

Provided are methods for determining the amount of total thiamine in a body fluid sample using liquid chromatography and mass spectrometry. Total thiamine is converted to free thiamine by treatment with an acid phosphatase prior to thiamine separation and quantification. 1. A method for determining the amount of total thiamine in a plasma or serum sample , comprising:(i) removing soluble protein from a plasma or serum sample;(ii) incubating the sample from step (i) with an acid phosphatase, for not longer than about 2 hours, to convert phosphorylated thiamine to thiamine;(iii) performing an organic solvent extraction of said sample from step (ii), wherein the result of said extraction is an organic solvent phase and an aqueous phase;(iv) purifying said thiamine from said aqueous phase of step (iii) by liquid chromatography; and(v) determining the amount of thiamine by mass spectrometry, wherein the amount of total thiamine in said sample is determined.2. The method of claim 1 , wherein the incubation of step (ii) is carried out at a pH of about 4.6±0.1.3. The method of claim 1 , wherein said acid phosphatase is carried out at a temperature of about 40° C.4. The method of claim 1 , wherein said incubation is carried out between about 1 and about 2 hours.5. The method of claim 1 , wherein soluble protein is removed in step (i) by treating said sample with an acid.6. The method of claim 1 , wherein said liquid chromatography comprises high performance liquid chromatography (HPLC).7. The method of claim 1 , wherein step (v) comprises ionizing said thiamine to a parent ion having a mass/charge ratio of 265.00±1.0.8. The method of claim 7 , wherein said parent ion is fragmented into one or more daughter ions and wherein the amount of one or more of said daughter ions is determined.9. The method of claim 8 , wherein said one or more daughter ions comprises an ion having a mass/charge ratio of 144.00±1.0 or 121.94±1.0.10. The method of claim 8 , wherein said one or more ...

METHODS FOR DETERMINATION OF PROTEIN PHOSPHATASE ACTIVITY, AND USES IN PREDICTING THERAPEUTIC OUTCOMES

One aspect of the present disclosure encompasses methods for determining a protein kinase or phosphatase activity in a biological sample, comprising: contacting in a reaction mix a first test sample and a fluorescently-labeled peptide substrate capable of being modified by a protein phosphatase or a protein kinase, contacting the reaction mix with a TiOmatrix, thereby partitioning fluorescently-labeled phosphorylated peptide from fluorescently-labeled dephosphorylated peptide; and determining the fluorescence of the fluorescently-labeled dephosphorylated peptide, thereby determining a protein kinase or phosphatase activity. 1. A method of determining calcineurin activity comprising the steps of:i) contacting in a reaction mix a first test sample and a fluorescently-labeled phosphorylated peptide substrate capable of being dephosphorylated by calcineurin, under conditions such that calcineurin dephosphorylates the fluorescently-labeled phosphorylated peptide; and{'sub': '2', 'ii) contacting the reaction mix with a TiOmatrix, thereby partitioning fluorescently-labeled phosphorylated peptide from fluorescently-labeled non-phosphorylated peptide providing a partition with fluorescently-labeled non-phosphorylated peptide; and'}iii) determining calcineurin activity wherein measuring an amount of fluorescence emitted by the partition with fluorescently-labeled non-phosphorylated peptide is an indication of calcineurin activity.2. The method of claim 1 , wherein the fluorescently labeled phosphorylated peptide has an amino acid sequence selected from SEQ ID NO.: 1 and SEQ ID NO.: 2.3. The method of claim 1 , wherein the fluorescently labeled phosphorylated peptide is capable of distinguishing a first isoform of calcineurin from a second isoform.4. The method of claim 2 , wherein the fluorescently labeled phosphorylated peptide has the amino acid sequence according to SEQ ID NO.: 1 claim 2 , is phosphorylated on the Ser-15 position claim 2 , and further comprises an N- ...

Methods for Diagnosis, Prognosis and Methods of Treatment

This invention is directed to methods and compositions for diagnosis, prognosis and for determining methods of treatment. The physiological status of a cell present in a sample (e.g. clinical sample) can be used in diagnosis or prognosis of a condition (e.g. Chronic Lymphocytic Leukemia), in patient selection for therapy, to monitor treatment and to modify or optimize therapeutic regimens. 1. A method for classifying a cell comprisingcontacting said cell with a modulator, wherein the modulator is a tyrosine phosphatase inhibitor;determining the presence or absence of a change in activation level of an activatable element in said cell; andclassifying said cell based on said presence or absence of said change in the activation level of said activatable element.2. The method of claim 1 , wherein said tyrosine phosphatase inhibitor is CD45-associated protein tyrosine phosphatase inhibitor claim 1 , PHPS1 claim 1 , PP1 claim 1 , PP2 claim 1 , Bay U6751 claim 1 , BVT.948 claim 1 , NSC 295642 claim 1 , PRL-3 Inhibitor I claim 1 , Phenylarsine oxide claim 1 , Sodium Stibogluconate claim 1 , Sodium orthovanadate claim 1 , pervanadate claim 1 , bisperoxovanadium claim 1 , phenylarsine oxide claim 1 , alendronate claim 1 , etidronate claim 1 , vanadate claim 1 , gallium nitrate claim 1 , suramin claim 1 , or aplidin.3. The method of claim 1 , wherein said tyrosine phosphatase inhibitor is hydrogen peroxide (HO).4. The method of wherein said change in activation level of an activatable element is an increase in activation level of an activatable element.5. The method of wherein said cell is a cancer cell.6. The method of wherein said presence or absence of a change in activation level of said activatable element is compared to a normal cell contacted with said modulator.7. The method of wherein said cell is a hematopoietically derived cell.8. The method of wherein the presence or absence of a change in the activation levels of a plurality of activatable elements is determined ...

Methods and Systems for Preparing Irreversible Inhibitors of Protein Tyrosine Phosphatases

Described herein are the preparation and use of novel bromo-phosphonomethylphenylalanine amino acid derivatives (BrPmp) and BrPmp-containing peptides as specific, irreversible protein tyrosine phosphatase inhibitors, which are suitable for application in peptide synthesis. These derivatives are particularly advantageous since their synthesis is both easy and scalable, and they are suitable for peptide synthesis. The BrPmp derivatives described herein can be appropriately protected to allow for solid phase peptide synthesis (SPPS) and incorporation into peptides for preparation of protein tyrosine phosphatase inhibitors and inhibitor libraries. The peptides and peptide libraries can be used to identify new protein tyrosine phosphatase specific sequences and profile protein tyrosine phosphatase activity in cell lysates, diagnostic samples and biopsy samples. 2. The bromo-phosphonomethylphenylalanine amino acid derivative of the Formula (I) of claim 1 , wherein R is hydrogen and each Ris hydrogen.3. The bromo-phosphonomethylphenylalanine amino acid derivative of the Formula (I) of claim 1 , wherein R is Fmoc claim 1 , Boc claim 1 , or Cbz and Ris methyl claim 1 , ethyl claim 1 , benzyl claim 1 , dimethylamino (—N(CH)) claim 1 , propylamino (—NHCHCHCH) claim 1 , isopropylamino (—NHCH(CH)) or allyl.423-. (canceled)24. The bromo-phosphonomethylphenylalanine amino acid derivative of the Formula (I) of claim 1 , which is an L-amino acid derivative.25. The bromo-phosphonomethylphenylalanine amino acid derivative of the Formula (I) of which is a D-amino acid derivative.2628-. (canceled)29. The bromo-phosphonomethylphenylalanine amino acid derivative of the Formula (II) of claim 1 , wherein Ris hydrogen claim 1 , Ris the side chain of aspartic acid claim 1 , nis 1 claim 1 , each Ris hydrogen claim 1 , Ris the side chain of leucine claim 1 , Ris hydroxyl claim 1 , and nis 1.30. The bromo-phosphonomethylphenylalanine amino acid derivative of the Formula (II) of claim 1 , wherein ...

In Situ Chemiluminescent Substrates and Assays

Methods for generating a chemiluminescent enzyme substrate in situ, in aqueous or other assay conditions. Also disclosed are methods to use the substrates to generate light, detect and/or quantify enzymes, antigens, and/or nucleic acids. Kits relating to these methods are also disclosed. 2. The method of claim 1 , wherein the oxidant is selected from hydrogen peroxide claim 1 , sodium molybdate claim 1 , hydrogen peroxide and sodium molybdate claim 1 , hypochlorite claim 1 , hypochlorite and hydrogen peroxide claim 1 , aryl endoperoxide claim 1 , calcium peroxide peroxyhydrate claim 1 , and combinations thereof.34.-. (canceled)6. The method of claim 1 , wherein Ris alkyl containing 1 to 2 carbon atoms or trifluoalkyl containing 1 to 2 carbon atoms.12. (canceled)13. The method of claim 1 , wherein the enzyme is a hydrolyic enzyme selected from alkaline phosphatase claim 1 , β-galactosidase claim 1 , β-glucosidase claim 1 , β-glucuronidase claim 1 , and neuraminidase.14. (canceled)15. The method of claim 1 , further comprising the step of detecting the light emitted from the reaction mixture after addition of the aqueous solution of the 1 claim 1 ,2-dioxetane enzyme substrate claim 1 , wherein the emission of light is indicative of the presence of the enzyme claim 1 , and the amount of light emitted can be correlated to the amount of the enzyme present in the sample.16. The method of claim 1 , wherein the enzyme moiety is an enzyme-linked antibody comprising a first antibody capable of binding to an antigen and an enzyme; an enzyme-linked antigen comprising an antigen and an enzyme; or an enzyme-linked oligonucleotide comprising an oligonucleotide capable of hydridizing to a nucleic acid claim 1 , wherein the enzyme is capable of cleaving the 1 claim 1 ,2-dioxetane enzyme substrate so that the substrate decomposes and generates light.17. (canceled)18. The method of claim 16 , wherein the first antibody or antigen is covalently linked to a label and the enzyme is ...

Method for Diagnosis of Bladder Cancer and Related Kits

The invention refers to a novel molecular biomarker, namely PTPD1, that is markedly increased in human bladder cancers. PTPD1 expression positively correlated with the grading and invasiveness potential of these tumors. PTPD1 can be detected at high levels in exfoliated bladder cells isolated from urine of bladder cancer patients, while no PTPD1 signal was evident in normal exfoliated bladder cells. Thus, PTPD1 detection in urine samples may represent a novel and reliable marker for non-invasive diagnosis of aggressive bladder cancer. 116-. (canceled)17. A method for diagnosing , or the monitoring control for therapy of , bladder cancer , said method comprising detecting (i) PTPD1 protein or an immunological fragment thereof , (ii) PTPD1 enzymatic activity , or (iii) PTPD1 mRNA in a body sample.18. The method of claim 17 , wherein the detecting of the PTPD1 protein or of an immunological fragment thereof comprises allowing the body sample to react with a PTPD1 protein specific ligand to form a complex; and detecting the complex.19. The method of claim 18 , wherein the PTPD1 protein specific ligand comprises an anti-PTPD1 antibody or a derivative thereof.20. The method of claim 19 , wherein the anti-PTPD1 antibody or a derivative thereof is obtained by using as an immunogen the whole PTPD1 protein of SEQ ID No. 1 or an immunogenic fragment thereof.2120. The method of claim of wherein the immunogenic fragment of the PTPD1 protein of SEQ ID No. 1 comprises between aa 751 and aa 910 of SEQ ID No. 1.22. The method of claim 21 , wherein the immunogenic fragment of the PTPD1 protein of SEQ ID No. 1 is a sequence between aa 751 and aa 910 of SEQ ID No. 1.23. The method of claim 18 , wherein the detecting step comprises detection of a specific fluorescent signal.24. The method of claim 17 , wherein the detecting of the PTPD1 enzymatic activity comprises performance by fluorescent or radiolabeled assays.25. The method of claim 17 , wherein the detecting of the PTPD1 mRNA ...

METHOD FOR LARGE SCALE PREPARATION OF THE ACTIVE DOMAIN OF HUMAN PROTEIN TYROSINE PHOSPHATASE WITHOUT FUSION PROTEIN

The present invention relates to methods for identifying inhibitors or activators of protein tyrosine phosphatase (PTP). In some examples, the methods utilize a PTP active domain with high activity and stability expressed without help of a fusion protein, by using computer based protein structure prediction technique. PTP prepared by the disclosed method may also be used as an antigen protein for the construction of a selective antibody and as a protein for the studies of PTP structure and functions. 1. A method for screening for a protein tyrosine phosphatase (PTP) activity inhibitor or activator in vitro , comprising the following steps: i) investigating homology among subgroups of PTP and selecting a region exhibiting high homology;', 'ii) examining whether the selected region of step i) corresponds to an active domain of a standard protein whose secondary and tertiary structures have already been identified;', 'iii) analyzing the secondary structure of the selected region of step i) if it corresponds to the active domain and then determining a boundary of PTP active domain by the location not containing helix or sheet of the secondary structure;', 'iv) determining 2-3 amino acids of the boundary of N-terminal and C-terminal of the PTP active domain primarily determined in step iii) to be a small amino acid or a charged amino acid by amino acid analysis;', 'v) constructing an expression vector containing a polynucleotide encoding the amino acids included in the inside of the boundary of the PTP active domain determined in step iv);', 'vi) generating a transformant by introducing the expression vector of step v) into a host cell; and,', 'vii) inducing expression of the recombinant PTP active domain by culturing the transformant of step vi) and obtaining the recombinant PTP active domain produced therefrom;, 'a) preparing a recombinant PTP active domain byb) contacting a PTP specific substrate and a candidate inhibitor or activator with the recombinant PTP active ...

METHODS FOR DETECTION OF RARE SUBPOPULATIONS OF CELLS AND HIGHLY PURIFIED COMPOSITIONS OF CELLS

Methods are provided for detection of a target cell type within a cell population, and compositions are provided comprising cells and an indicator that indicates the number of cells of the target cell type in the cell population. Examples are provided in which these methods are used to detect human embryonic stem cells within a differentiated cell population with exquisite sensitivity. Differentiated cells produced from embryonic stem cells can be characterized by these methods before transplantation into a recipient, thereby providing further assurance of safety. 1. A method of detecting the presence of or confirming the absence of target cells in a cell population , comprising:(a) providing a cell population;(b) applying a first stain and a second stain to said cell population, wherein said first stain detects a first marker the expression of which is indicative of the presence of target cells and said second stain detects a second marker the expression of which is also indicative of the presence of the same target cells, and wherein said first stain is detectable under visible light and said second stain is detectable under ultraviolet light;(c) microscopically observing cells of said cell population under visible light to detect any cells that are positive for said first marker;(d) microscopically observing said cells that are positive for said first marker under ultraviolet light and determining whether any of said detected cells are positive for said first marker and said second marker, and(e) identifying any cells that are positive for said first marker and said second marker as target cells.2. A method of detecting the presence of or confirming the absence of target cells in a cell population , comprising:(a) providing a cell population;(b) applying a first stain and a second stain to said cell population, wherein said first stain detects a first marker the expression of which is indicative of target cells and said second stain detects a second marker the ...

SUBSTRATES AND METHODS FOR STAINING LIVE STEM CELLS

The invention relates to novel substrates and methods for staining live stem cells. The stain may be used to identify induced pluripotent stem cell colonies during the process of somatic cell reprogramming. 1. A composition comprising a substrate capable of identifying a live stem cell of interest , wherein the substrate is cell permeable and is capable of being modified by the stem cell into a modified substrate , wherein the modified substrate is both permeable and detectable such that the live stem cell can be identified by detecting the presence of the modified substrate , and wherein the substrate and the modified substrate are non-toxic.2. The composition of wherein the live stem cell of interest is selected from the group consisting of an embryonic stem cell (ESC) claim 1 , an induced pluripotent stem cell (iPSC) claim 1 , a pluripotent cell claim 1 , a progenitor cell claim 1 , a reprogrammed cell claim 1 , and a dedifferentiated cell.3. The composition of wherein the detecting is of a fluorescent signal.4. The composition according to claim 1 , wherein the substrate is chosen from the group consisting of xanthene derivatives claim 1 , coumarin derivatives claim 1 , resorufin derivative claim 1 , triphenylmethanes and dialkylacridinones.5. The composition according to claim 1 , wherein the substrate is a rosamine derivative.6. The composition according to claim 4 , wherein the xanthene derivative is selected from the group consisting of fluorescein derivatives and rhodamine derivatives.7. The composition according to claim 6 , wherein the fluorescein derivative is either a fluorescein monophosphate or a fluorescein diphosphate.10. The composition according to claim 4 , wherein the coumarin derivative is an umbelliferone derivative.12. The composition according to claim 11 , wherein the compound is 6 claim 11 ,8-difluoro-4-methylumbelliferyl phosphate (DiFMUP).14. The composition according to claim 4 , wherein the substrate is reactive with enzymes chosen ...

SMALL MOLECULE ANTAGONISTS OF DUSP5 AND METHODS OF USE

The present invention is directed to compounds that specifically target DUSP5 and act as antagonists of that enzyme. Such compounds are useful in the treatment of various conditions including, but not limited to vascular anomalies, cancer, and macular degeneration. 1. A pharmaceutical composition comprising a compound , or a pharmaceutically acceptable salt , solvate , or hydrate thereof , and a pharmaceutically acceptable carrier , wherein the compound comprises two negatively charged sulfonate groups or bioisosteric sulfonate analog groups tethered by a core scaffold of biphenyl or a planar analog of biphenyl , and wherein the core scaffold separates the negatively charged groups at a distance of about 6 to about 8 Angstroms.2. The composition of claim 1 , wherein the core scaffold comprises one or more of fluorene claim 1 , biphenyl claim 1 , carbazole claim 1 , dibenzofuran claim 1 , calixarene claim 1 , and naphthalene.4. The composition of claim 1 , wherein the negatively charged sulfonate groups or bioisosteric sulfonate analog groups are separated by a distance of about 7 angstroms.5. The composition of claim 1 , wherein the bioisosteric sulfonate analog groups are selected from the group consisting of sulfonamide claim 1 , tetrazole claim 1 , and carboxylic acid.6. The composition according to claim 1 , wherein said compound is a dual specificity phosphatase-5 (DUSP5) inhibitor.7. A medicament for treating claim 1 , preventing claim 1 , or alleviating a vascular anomaly comprising at least one compound as defined in in an effective amount to antagonize DUSP5.8. A method of treating claim 1 , preventing claim 1 , or alleviating a vascular anomaly comprising administering to a subject in need thereof a therapeutically effective amount of a compound or composition defined in wherein the compound antagonizes dual specificity phosphatase-5 (DUSP5).9. The method of claim 7 , wherein the vascular anomaly is associated with a Serine to Proline mutation at amino ...

METHODS TO ASSAY PHOSPHATASE ACTIVITY

Methods and kits for enzymes involved in post-translational modifications are provided. The methods employ elemental analysis, including ICP-MS. The methods allow for the convenient and accurate analysis of post-translation modifications of substrates by enzymes involved in post-translational modifications, including kinase and phosphatase enyzmes 159-. (canceled)60. A method for a phosphatase assay , comprising:a) incubating free phosphorylated substrate labeled with an element tag with a support having attached thereto metal ion coordination complexes;b) separating free phosphorylated substrate from bound phosphorylated substrate labeled with an element tag attached to the metal ion coordination complexes attached to the support;c) incubating ADP and at least one phosphatase with the bound phosphorylated substrate labeled with an element tag attached to the metal ion coordination complexes attached to the support under conditions to enable the phosphatase to dephosphorylate the substrate;d) separating free non-phosphorylated substrate labeled with an element tag from bound phosphorylated substrate attached to the metal ion coordination complexes attached to the support; ande) using elemental analysis to detect the element tag associated with the free non-phosphorylated substrate.61. The method of wherein a multitude of free phosphorylated substrates each labeled with an element tag that is characteristic of the substrate are incubated in step (a) and the tag elements characteristic of the substrate are measured in step (e).62. The method of where in step (c) the conditions to enable the phosphatase to dephosphorylate the substrate include incubation with a phosphatase reaction buffer.63. The method of wherein step (c) comprises incubating antagonists or agonists of phosphatase.64. The method of wherein the metal ion coordination complex is attached to element labeled beads.65. The method of wherein the metal ion coordination complex attached to the support is a ...

ISOLATION OF ADULT MULTIPOTENTIAL CELLS BY TISSUE NON-SPECIFIC ALKALINE PHOSPHATASE

The present invention relates to the use of tissue non-specific alkaline phosphatase (TNAP) as a marker for identifying and/or isolating adult multipotential cells. The present invention also relates to cell populations enriched by methods of the present invention and therapeutic uses of these cells. 145-. (canceled)46. An enriched population of TNAP adult multipotential cells , wherein at least 4% of the total cell population are TNAP , STRO-1 multipotential cells capable of being cultured in vitro to produce adipocytes , osteocytes and chondrocytes , wherein the TNAP marker is a TNAP marker which binds the STRO-3 antibody produced by the hybridoma cell line deposited under ATCC Accession Number PTA-5582.47. An enriched population according to claim 46 , wherein at least 10% of the total cell population are TNAP claim 46 , STRO-1 multipotential cells capable of being cultured in vitro to produce adipocytes claim 46 , osteocytes and chondrocytes.48. An enriched population according to claim 46 , wherein at least 20% of the total cell population are TNAP claim 46 , STRO-1 multipotential cells capable of being cultured in vitro to produce adipocytes claim 46 , osteocytes and chondrocytes.49. An enriched population according to claim 46 , wherein the STRO-1 cells are STRO-1.50. An enriched population according to claim 47 , wherein the STRO-1 cells are STRO-1.51. An enriched population according to claim 48 , wherein the STRO-1 cells are STRO-1.52. A method of generating a committed cell population of a specific tissue type claim 46 , the method comprising culturing a population of adult multipotential cells according to in the presence of one or more stimulatory factors; and subjecting said cultured population to conditions biasing differentiation of the adult muitipotential cells to the specific tissue type.53. A method of generating a committed cell population of a specific tissue type claim 47 , the method comprising culturing a population of adult multipotential ...

Compounds for Treating Tauopathies and Restless Leg Syndrome and Methods of Using and Screening for Same

Provided are compounds capable of enhancing the ability of receptor-type tyrosine-protein phosphatase delta (PTPRD) to dephosphorylate a kinase. Also disclosed is a method of treating a tauopathy or restless leg syndrome in a subject comprising administering to the subject an effective amount of a disclosed compound. Also disclosed are kits comprising the compounds together with instructions for treating a condition and/or a compound known for treating the condition. Finally, disclosed herein is a screening method suitable for identifying positive allosteric modulators of the ability of a receptor-type tyrosine-protein phosphatase delta (PTPRD) to dephosphorylate a kinase. 3. The method of claim 1 , wherein the compound is Quercetin.4. The method of claim 1 , wherein the tauopathy is Alzheimer's disease claim 1 , chronic traumatic encephalopathy claim 1 , corticobasal degeneration claim 1 , frontotemporal lobar degeneration claim 1 , behavioral variant frontotemporal dementia claim 1 , language variant frontotemporal dementia claim 1 , right temporal variant frontotemporal dementia claim 1 , Pick disease claim 1 , or progressive supranuclear palsy.5. The method of claim 1 , further comprising administering to the subject an effective amount of a compound known for treating the tauopathy or restless leg syndrome.8. The method of claim 6 , wherein the compound is Quercetin.9. The method of claim 6 , wherein the kinase is glycogen synthase kinase GSK3β claim 6 , glycogen synthase kinase GSK3α claim 6 , cyclin dependent kinase-5 CDK5 claim 6 , or a combination thereof.12. The kit of claim 10 , wherein the compound represented by Formula (I) is Quercetin.13. The kit of claim 10 , wherein the tauopathy is Alzheimer's disease claim 10 , chronic traumatic encephalopathy claim 10 , corticobasal degeneration claim 10 , frontotemporal lobar degeneration claim 10 , behavioral variant frontotemporal dementia claim 10 , language variant frontotemporal dementia claim 10 , right ...

Method of evaluating quality of dephosphorylation reagent and method of detecting target nucleic acid

A method evaluates a quality of a dephosphorylation reagent, the method including the steps of: providing a dephosphorylation reagent containing an alkaline phosphatase and a peptide fragment derived from the alkaline phosphatase; and evaluating the dephosphorylation reagent as having a high quality if a content ratio of the peptide fragment to the alkaline phosphatase is a predetermined reference value or less.

MEDIUM FOR DETECTING STAPHYLOCOCCUS AUREUS, SHEET FOR DETECTING S. AUREUS COMPRISING SAME, AND METHOD FOR DETECTING S. AUREUS USING SAME

The purpose of the present invention is to provide a detection means whereby can be identified at a high accuracy. An aspect of the present invention relates to a medium for detecting which comprises one or more kinds of nutrient components, a color developing agent capable of developing a color in the presence of α-glucosidase, a color developing agent capable of developing a color in the presence of phosphatase and 0.5 mg/cmor more of sodium colistin methanesulfonate. Another aspect of the present invention relates to a sheet for detecting , said sheet comprising the aforesaid medium, and a method for detecting with the use of the medium and sheet as described above. 16-. (canceled)7Staphylococcus aureusStaphylococcus aureus. A microorganism culture substrate used for detection of comprising a substrate and a culture layer provided on an upper surface of the substrate , wherein the culture layer comprises a medium used for detecting comprising one or more nutrient components , a color developer that develops color in the presence of α-glucosidase , a color developer that develops color in the presence of phosphatase , and colistin sodium methanesulfonate at 0.5 mg/cmor more.8. The microorganism culture substrate according to claim 7 , which further comprises the substrate in the form of a sheet and a cover sheet covering the culture layer claim 7 , wherein the culture layer further comprises polyvinylpyrrolidone and one or more gelling agents.9Staphylococcus aureus. A method for detecting comprising:{'claim-ref': {'@idref': 'CLM-00007', 'claim 7'}, 'a step of sample addition comprising adding a microorganism-containing sample to the medium or the culture layer of the microorganism culture substrate according to ;'}a step of colony formation comprising incubating the medium or microorganism culture substrate added with the sample to form a microbial colony; and{'i': 'Staphylococcus aureus', 'a step of strain identification comprising identifying on the basis of the ...

MEDICAL DEVICE DESIGN, MANUFACTURE AND TESTING SYSTEMS

Described are methods and systems for testing lumens of cannulated delivery components or assemblies thereof that may be used to deliver cells to a patient. Test cells are contacted with walls of the lumens and/or liquids that contact walls of the lumens, potentially over an incubation period. The test cells are then assessed for an effect of the wall contact, or the liquid contact, on at least one and preferably multiple characteristics of the test cells such as innate immune response, metabolic activity, viability, cytotoxic response, and/or motility. Methods and systems as described can be used in the development and/or manufacture of cannulated delivery devices, for example providing specifications for design or process inputs or outputs, design or process validations, and/or device lot approvals. Also described are devices or products produced in accordance with such methods and systems. 1. A method for testing a region of a medical device , comprising:incubating a liquid medium in contact with a wall of the region;contacting at least one cell with the liquid medium; andassessing the affect of said contacting on the expression of at least one toll-like receptor by the cell.2. The method of claim 1 , wherein the medical device is a medical delivery device claim 1 , and the region is a lumen.3. The method of claim 1 , wherein the at least one cell includes exogenous DNA encoding the at least one toll-like receptor.4. The method of claim 1 , wherein the cell is a mammalian cell.5. The method of claim 1 , wherein the cell is a human cell.69-. (canceled)10. The method of claim 1 , wherein the at least one toll-like receptor includes toll-like receptor 2 claim 1 , toll-like receptor 4 claim 1 , or both.1113-. (canceled)14. A method for testing a region of a medical device claim 1 , comprising:first assessing the region for an effect on innate immune response of a cell; andsecond assessing the region for an effect on at least one other characteristic of a cell, said ...

Detection and quantification of analytes based on signal induced by alkaline phosphate

A general methodology for highly sensitive and selective sensors that can achieve portable, low-cost and quantitative detection of a broad range of targets using only a personal glucose meter (PGM) is disclosed. The method and sensors take advantage of the ability of alkaline phosphatase (ALP) to convert glucose-1-phosphate to glucose, and the ability of PGMs to detect the generated glucose. The disclosed sensors can be part of a lateral flow device. Methods of using such sensors for detecting target agents, for example to diagnose disease, are also provided.

SYSTEMS AND METHODS FOR MULTI-ANALYSIS

Systems and methods are provided for sample processing. A device may be provided, capable of receiving the sample, and performing one or more of a sample preparation, sample assay, and detection step. The device may be capable of performing multiple assays. The device may comprise one or more modules that may be capable of performing one or more of a sample preparation, sample assay, and detection step. The device may be capable of performing the steps using a small volume of sample. 18-. (canceled)9. A sample processing system , comprising:one or more operational matrices, wherein at least one of said operational matrices comprises one or more operational states of the sample processing system, and further wherein at least one of said operational states comprises two or more sample processing routines to be sequentially performed by the sample processing system; a sample handling system comprising at least one pipette nozzle for engaging with at least one pipette tip;', 'at least one detector; and', 'a cartridge receiving location;, 'a sample processing device comprising at least one pipette tip;', 'at least one assay unit;', 'at least one reagent unit; and', 'at least one sample collection unit for receiving a sample., 'and the cartridge comprising10. The system of claim 9 , wherein the at least one sample collection unit is configured to receive a sample of no more than about 500 μl.11. A method for restoring an operational state of a sample processing system claim 9 , comprising:{'claim-ref': {'@idref': 'CLM-00009', 'claim 9'}, 'accessing a sample processing catalog of the sample processing system of following a fault condition of the sample processing system; wherein the sample processing catalog comprises a record of operational data of the one or more operational states of the sample processing system;'}identifying a last-in-time operational state of the sample processing system from the sample processing catalog;identifying a last-in-time sample processing ...

USE OF AT LEAST ONE CHROMOGENIC AND/OR FLUOROGENIC PHOSPHATASE SUBSTRATE FOR THE DETECTION AND/OR ENUMERATION OF ENTEROBACTERIA IN A SAMPLE

Use of at least one chromogenic and/or fluorogenic phosphatase substrate for the detection and/or enumeration of enterobacteria in a sample likely to contain them, such as a food sample. 1. Use of at least one chromogenic and/or fluorogenic phosphatase substrate for the detection and/or enumeration of enterobacteria in a sample likely to contain them , such as a food sample , said at least one chromogenic and/or fluorogenic phosphatase substrate being a synthetic substrate comprising two parts , a first part specific to phosphatase activity and a second chromogenic and/or fluorogenic marker part.2. Use according to claim 1 , wherein said phosphatase substrate is selected from:substrates based on indoxyl or a derivative thereof, such as 5-bromo-4-chloro-3-indoxyl-phosphate, 5-bromo-6-chloro-3-indoxyl-phosphate, 6-chloro-3-indoxyl-phosphate, 5-iodo-3-indoxyl-phosphate, 6-bromo-3-indoxyl-phosphate, 5,6-dibromo-3-indoxyl-phosphate, 5-bromo-3-indoxyl-phosphate, Aldol® 458 phosphate, Aldol® 470 phosphate, Aldol® 484 phosphate, Aldol®495 phosphate, Aldol®515 phosphate, Aldol® n phosphate, and3-hydroxyflavone-phosphate.3. Use according to or claim 1 , wherein said phosphatase substrate is a substrate based on indoxyl or a derivative thereof claim 1 , said phosphatase substrate being used in combination with at least one agent promoting oxidative polymerization of the indoxyl derivative claim 1 , such as an ammonium ferric citrate metal complex claim 1 , preferably said phosphatase substrate being chosen from 5-bromo-4-chloro-3-indoxyl-phosphate claim 1 , 5-bromo-6-chloro-3-indoxyl-phosphate claim 1 , 6-chloro-3-indoxyl-phosphate claim 1 , 5-iodo-3-indoxyl-phosphate claim 1 , 6-bromo-3-indoxyl-phosphate claim 1 , 5 claim 1 ,6-dibromo-3-indoxyl-phosphate claim 1 , 5-bromo-3-indoxyl-phosphate.4. Use according to one of to claim 1 , wherein said phosphatase substrate is contained within a composition comprising at least one claim 1 , preferably two claim 1 , and advantageously ...

Method for measuring calcineurin activity

The present invention relates to a method for measuring the activity of calcineurin in a biological sample wherein a kinase inhibitor is present in the assay reaction mix.

Methods for identifying modulators of ras using nonlinear techniques

Provided herein are compositions and methods for identifying and detecting modulators of Ras protein conformational states through the use of second harmonic generation (SHG) technology. Also provided herein are methods for detecting a conformational changes in the three dimensional structure of a protein bound to a supported lipid bilayer.

DATA COLLECTION METHOD TO BE USED FOR CLASSIFYING CANCER LIFE

Blood serum is applied on a support that has been impregnated in a buffer solution, the support is fractionated by electrophoresis at a predetermined liquid temperature to isolate proteins in the blood serum, an ALP isozyme is detected by color-developing with an ALP isozyme staining solution, the mobility, chromosome shape, density, and the like of each isozyme are determined by matching the protein fraction image against the ALP isozyme, development of a minute cancer that is occurring is discovered, and also the risk of tumor and risk of cancer are evaluated by matching against the analysis results of a tumor marker to classify the life of the cancer for early cancer discovery. 1. A data collection method to be used for classifying cancer life to discover development of minute cancer occurring in a human body and to perform data analysis of risk of tumor and risk of cancer in two divided stages of a preclinical cancer stage and a clinical cancer stage , wherein ,for the data used for analysis of a preclinical cancer stage,in order to collect data of an occurrence and a ratio of ALP I and activity value of ALP II and ALP III from patterns of the ALP I to the ALP IV as the ALP isozyme, examine the ratio of the numerical data, and perform data analysis for proliferation activity of cancer cells in view of APA calculated from ALP isozyme angle showing sharpness of the ALP II and the ALP III, andadditionally, for avoiding a cause contributed by an inflammatory disease, to perform an analysis while subtracting numerical data of a related part also occurring in the inflammatory disease from each numerical data obtained by measuring both the C reactive protein value (C inflammatory protein value, CRP value) and sialic acid value so as to perform data analysis for the existence and proliferation status of occurring minute cancer, andfor the data used for analysis of a clinical cancer stage,to collect data from a changed state like a decrease in albumin fraction and an ...

Flash and Glow 1,2-Dioxetanes

Compounds having chemiluminescent flash and glow properties. Also disclosed are methods using the compounds to generate light, detect and/or quantify enzymes, antigens, and/or nucleic acids. Also disclosed are kits relating to these compounds. 4. The compound of claim 1 , wherein Ris a straight chain alkyl comprising 1 to 5 carbon atoms which may be optionally substituted with one or more halogens atoms.5. The compound of claim 1 , wherein Ris a straight chain alkyl comprising 1 to 2 carbon atoms or straight chain trifluoalkyl comprising 1 to 2 carbon atoms.6. The compound of claim 5 , wherein Ris methyl or 1 claim 5 ,1 claim 5 ,1-trifluoroethyl.13. The compound of claim 1 , wherein the electron-donating group is a straight chain alkyl comprising 1 to 20 carbon atoms claim 1 , a branched alkyl comprising 3 to 20 carbon atoms claim 1 , a straight chain alkoxy comprising 1 to 20 carbon atoms claim 1 , or a branched alkoxy comprising 3 to 20 carbon atoms.14. The compound of claim 1 , wherein the electron-donating group is a straight chain alkyl comprising 1 to 20 carbon atoms or straight chain alkoxy comprising 1 to 20 carbon atoms.15. The compound of claim 1 , wherein the electron-donating group is a straight chain alkyl comprising 1 to 5 carbon atoms or straight chain alkoxy comprising 1 to 5 carbon atoms.16. The compound of claim 1 , wherein the electron-donating group is selected from the group consisting of methyl claim 1 , ethyl claim 1 , propyl claim 1 , butyl claim 1 , methoxy claim 1 , ethoxy claim 1 , propyloxy and butyloxy.17. The compound of claim 1 , wherein the cleavage of the bond cleavable by an enzyme moiety results in the generation of light at 25° C. which reaches a maximum in less than about 15 minutes.18. The compound of claim 17 , wherein the generation of light reaches a maximum in less than about 10 minutes.19. The compound of claim 17 , wherein the generation of light reaches a maximum in less than about 5 minutes.20. The compound of claim 1 , ...

SENSOR FOR DETECTING STEM CELL DIFFERENTIATION BASED ON ELECTROCHEMICAL METHODS

This invention relates to a sensor for detecting a stem cell differentiation, including (a) an electrode; and (b) a substrate of an alkaline phosphatase. The phosphorylation or dephosphorylation of the substrate for an alkaline phosphatase as a stem cell undifferentiation marker which dephosphorylates its substrate may be measured using an electrical signal in the present sensor. Therefore, the sensor of the present invention enables to electrically detect a stem cell status in a high-throughput manner and to determine the stem cell differentiation. 1(a) preparing the sensor for detecting the stem cell differentiation, comprising (i) an electrode which comprises gold electrode as the working electrode, a platinum electrode as the counter electrode, and a Ag/AgCl electrode as the reference electrode;and (ii) 1-Naphthyl phosphate (1-NP), a substrate for an alkaline phosphatase;(b) contacting cells of interest to the sensor; and(c) measuring the phosphorylation or dephosphorylation of 1-NP for alkaline phosphatase on the cells through an electrical signal, wherein the stem cell comprises an alkaline phosphatase where the stem cell is an undifferentiated stem cell; and the stem cell comprises no alkaline phosphatase where the stem cell is a differentiated stem cell.. A method for determining a stem cell differentiation, comprising the steps of: This is a continuation application of Application No. 12/579,084 filed on Oct. 14, 2009, which claims under 35 U.S.C. §119(a) the benefit of Korean Application No. 10-2009-0026353 filed Mar. 27, 2009, which applications are incorporated herein by reference.1. Technical FieldThe present invention relates to a sensor for detecting a stem cell differentiation, including: (a) an electrode; and (b) a substrate for an alkaline phosphatase.2. Background ArtCell chip technology is a promising tool for utilization in cell based assays. There are two kinds of cell detection systems for these chips. These systems are based on optical ...

NANOPATTERNED EXTRACELLULAR MATRICES ENABLE CELL-BASED ASSAYS WITH A MASS SPECTROMETRIC READOUT

The present disclosure provides methods in which adherent cells are treated with small molecules, cultured, lysed, and then analyzed by mass spectrometry to measure the activities of endogenous enzymes. The implementation of this method relies on the use of surfaces that are nanopatterned with cell adhesion ligands to mediate cell attachment and a peptide that is a substrate for the desired enzyme activity in the lysate. 1. A method of assaying activity of an intracellular enzyme , comprising: (i) coating a polymer pen lithography (PPL) tip array with a first monolayer reagent and printing the first monolayer reagent at selected positions on the surface to form an array of printed first monolayer reagent,', '(ii) incubating the array of printed first monolayer reagent with a second monolayer reagent such that the second monolayer reagent is adsorbed onto unprinted portions of the surface,, '(a) printing a surface with an array of immobilized cell adhesion ligands and immobilized substrates for the intracellular enzyme by'} (iii) contacting the resulting array of step (ii) with the substrate for the intracellular enzyme under conditions to immobilize the substrate to the surface at the portion of the surface comprising the monolayer reagent for chemical immobilization,', '(iv) contacting the resulting array of step (ii) with the cell adhesion ligand under conditions to immobilize the cell adhesion ligand to the surface at the portion of the surface comprising the monolayer reagent for adsorption of the cell adhesion ligand; wherein steps (iii) and (iv) can be performed in either order;, 'wherein one of the first monolayer reagent and the second monolayer reagent comprises a monolayer reagent for adsorption of the cell adhesion ligand and the other comprises a monolayer reagent for chemical immobilization of the substrate for the intracellular enzyme,'}(b) contacting a cell and the surface of step (a), the contacting resulting in immobilization of the cell via ...

Method and Device for Generating a Tunable Array of Fluid Gradients

Provided herein are devices and methods for generating microfluidic gradients, including an array of unique microfluidic gradients within an array of microchannels. Fluids within conduits are mixed in an intersection region to generate a mixed flow stream in a source reservoir channel that provides a gradient that varies with axial distance from the intersection region. Microchannels having an inlet connected to the source reservoir channel are configured to provide a microfluidic gradient in the microchannel. An outlet end of the microchannel is connected to a sink reservoir channel. By varying the ratio of fluid flow rates from the fluid conduits, the microchannel gradients are tuned. In this manner, a large number of unique gradients or array of microfluidic gradients is provided, wherein the gradient can be any number of physical or chemical parameters, including concentrations and physical fluid properties. 1. A microfluidic gradient generator for tuning dynamic components of fluid comprising:a first fluid conduit;a second fluid conduit,an intersection region that fluidically connects the first fluid conduit and the second fluid conduit, the intersection region comprising an intersection opening between the first fluid conduit and the second fluid conduit and a flow-divider that extends in a downstream direction from the intersection opening;a source reservoir channel fluidically connected to the intersection region and extending downstream from the intersection opening;a sink reservoir channel fluidically connected to the intersection region and extending downstream from the intersection opening;a microchannel array comprising a plurality of microchannels, each microchannel having an inlet end connected to the source reservoir channel and an outlet end connected to the sink reservoir channel, wherein adjacent microchannels are separated from each other by a separation distance, wherein the microchannel array traverses an axial distance along the source ...

METHODS FOR SCREENING VOLTAGE-GATED PROTEINS

In one aspect, the invention relates to a method for identifying a compound which modulates the activity of a voltage-gated protein. In certain embodiments, the voltage gate protein is a voltage-gated ion channel. In certain embodiments, the voltage-gated protein is a voltage sensitive phosphatase. In certain embodiments, the voltage-gated protein used in conjunction with the methods of the invention is modified to altered permeability or voltage sensitivity. 1. A method for identifying a compound which modulates the activity of a voltage-gated ion channel , the method comprising(a) providing a voltage-gated ion channel in a structure that separates a first medium from a second medium, wherein the voltage-gated ion channel exhibits independent ion permeation,(b) contacting the voltage-gated ion channel with a test compound,(c) measuring the amount of said independent ion permeation through the voltage-gated ion channel between the first and second media, and(d) comparing the amount of said independent ion permeation measured for the voltage-gated ion channel contacted with the test compound to the amount of said independent ion permeation measured for the voltage-gated ion channel not contacted with the test compound, wherein an increase or decrease in the amount of said independent ion permeation of the voltage-gated ion channel contacted with the test compound compared to the voltage-gated ion channel not contacted with the test compound indicates that the test compound modulates the activity of the voltage-gated ion channel.2. A method for identifying a compound which modulates the activity of a voltage-sensitive phosphatase , the method comprising(a) providing a voltage-sensitive phosphatase in a structure that separates a first medium from a second medium,(b) contacting the voltage-sensitive phosphatase with a test compound,(c) measuring the activity of the voltage-sensitive phosphatase, and(d) comparing the amount of said activity measured for the voltage- ...

DEGRADABLE CARBON NANOTUBE-CONTAINING BIOSENSORS AND METHODS FOR TARGET CLINICAL MARKER DETECTION

The invention relates to carbon nanotube-containing composites as biosensors to detect the presence of target clinical markers, methods of their preparation and uses in the medical field. The invention is particularly suitable for the detection in patient biological specimens of bone markers and tissue markers. The biosensors of the invention include carbon nanotubes deposited on a substrate, gold nanoparticles deposited on the carbon nanotubes and, binder material and biomolecule deposited on the gold-coated carbon nanotubes. The biomolecule is selected to interact with the target clinical markers. The biosensor can be used as an in-situ or an ex-situ device to detect and measure the presence of the target clinical markers. 1. An ex-situ , biosensor to detect a target clinical marker in a biological fluid sample of a patient , comprising:a substrate;a plurality of carbon nanotubes deposited on the substrate;a plurality of gold nanoparticles electrodeposited on the plurality of carbon nanotubes;a binding material adsorbed on the plurality of gold nanoparticles;a biotinylated biomolecule selected from the group consisting of antibody and aptamer, deposited on the binding material;a target clinical marker selected from the group consisting of a bone marker and a tissue marker,wherein the biotinylated biomolecule binds with the binding material, and interacts with the target clinical marker in the biological fluid sample of the patient; anda mechanism to provide an indication of the presence or absence of the target clinical marker in the biological fluid sample of the patient.2. The biosensor of claim 1 , wherein the target clinical marker is selected from the group consisting of c-terminal telopeptide claim 1 , n-terminal telopeptide claim 1 , alkaline phosphatase claim 1 , Troponin I and myoglobin.3. The biosensor of claim 1 , wherein the biotinylated biomolecule is selected from the group consisting of c-terminal telopeptide antibody claim 1 , n-terminal ...

SYSTEMS AND METHODS FOR MULTI-ANALYSIS

Systems and methods are provided for sample processing. A device may be provided, capable of receiving the sample, and performing one or more of a sample preparation, sample assay, and detection step. The device may be capable of performing multiple assays. The device may comprise one or more modules that may be capable of performing one or more of a sample preparation, sample assay, and detection step. The device may be capable of performing the steps using a small volume of sample. 1. A biological sample processing device comprising:a) a sample handling system;b) a detection station;c) a cytometry station comprising an imaging device and a stage for receiving a microscopy cuvette, andd) an assay station configured to support multiple components comprising i) a biological sample and ii) at least a first, a second, and a third fluidically isolated assay unit,wherein the sample handling system is configured to i) transfer at least a portion of the biological sample to the first assay unit, and the second assay unit, and the third assay unit; ii) transfer the first and second assay units containing biological sample to the detection station; and iii) transfer the third assay unit containing biological sample to the cytometry station.28.-. (canceled) This application is a continuation-in-part application of PCT Application No. PCT US2012/057155 and is a continuation-in-part of U.S. patent application Ser. No. 13/244,952 which claims priority to PCT Application No. PCT/US2011/53188, filed Sep. 25, 2011. All of the foregoing applications are incorporated herein by reference in their entirety for all purposes.The majority of clinical decisions are based on laboratory and health test data, yet the methods and infrastructure for collecting such data severely limit the quality and utility of the data itself. Almost all errors in laboratory testing are associated with human or pre-analytic processing errors, and the testing process can take days to weeks to complete. Often ...

COMPOUNDS FOR TREATMENT OF INTRACTABLE EPILEPSY AND DOORS SYNDROME

The present invention relates to the field of neurodegenerative diseases, and the prevention and/or treatment of TBC1D24-associated disorders, such as DOORS syndrome, (intractable) epilepsy, and nonsyndromic deafness. In particular, the present invention relates to phosphoinositide phosphatase inhibitors and screening methods for producing such inhibitors, more particularly, Synaptojanin-1 5′ phosphatase inhibitors, to increase phosphatidylinositol 4,5-bisphosphate (PI(4,5)P) levels in neuronal cells. 1. A method of treating a TBC1D24-associated disorder in a subject , the method comprising:{'sub': '2', 'treating the subject with a phosphoinositide phosphatase inhibitor which increases PI(4,5)Plevels in a cell.'}2. The method according to claim 1 , wherein the TBC1D24-associated disorder is selected from the group consisting of severe forms of epilepsy claim 1 , early onset epilepsy claim 1 , familial infantile myoclonic epilepsy (FIME) claim 1 , focal epilepsy claim 1 , dysarthria claim 1 , myoclonic epilepsy with dystonia claim 1 , familial malignant migrating partial seizures of infancy claim 1 , De Flippo malignant migrating partial seizures of infancy claim 1 , variable degrees of intellectual disability claim 1 , hearing loss claim 1 , nonsyndromic deafness claim 1 , autosomal-dominant nonsyndromic hearing loss claim 1 , and dominant nonsyndromic hearing impairment claim 1 , cortical myoclonus and cerebellar ataxia claim 1 , and DOORS syndrome (deafness claim 1 , onychodystrophy claim 1 , osteodystrophy claim 1 , mental retardation and seizures).3. The method according to claim 1 , wherein the phosphoinositide phosphatase is Synaptojanin-1.4. The method according to claim 3 , wherein the phosphoinositide phosphatase is the Synaptojanin-1 5′ phosphatase.5. The method according to claim 1 , wherein the cell is a neuronal cell.6. The method according to claim 1 , wherein the phosphoinositide phosphatase inhibitor is selected from the group consisting of bpV(pic) ...

COMPARTMENTALISED SCREENING BY MICROFLUIDIC CONTROL

The invention describes a method for the identification of compounds which bind to a target component of a biochemical system or modulate the activity of the target, comprising the steps of: a) compartmentalising the compounds into microcapsules together with the target, such that only a subset of the repertoire is represented in multiple copies in any one microcapsule; and b) identifying the compound which binds to or modulates the activity of the target; wherein at least one step is performed under microfluidic control. The invention enables the screening of large repertoires of molecules which can serve as leads for drug development. 1102-. (canceled)103. A method for screening a compound or compounds capable of modulating the activity of a target , comprising the steps of:(a) compartmentalising the compounds into microcapsules, such that only a subset of the repertoire is represented in multiple copies in any one microcapsule;(b) contacting a target having a desired activity with the compound or compounds and monitoring the modulation of an activity of the target by the compound or compounds, wherein one or more of steps a, b and c is performed under microfluidic control; and(c) identifying and sorting the microcapsules which contain the compound(s) having the desired activity using a change in their optical properties.104115-. (canceled)116. The method of claim 103 , wherein the target is compartmentalized together with the compounds in the microcapsules.117. The method of claim 103 , wherein a fluid stream comprising the target is introduced into the microcapsules comprising the compounds.118. The method of claim 103 , further comprising fusing a microcapsule comprising the target with each of the microcapsules comprising the compounds.119. The method of claim 103 , further comprising attaching the repertoire of compounds to microbeads.120. The method of claim 119 , wherein each microbead comprises a detectable tag.121. The method of claim 119 , wherein the ...

ULTRA-HIGH-SENSITIVE ASSAY OF PROTEIN AND NUCLEIC ACID AND KIT, AND NOVEL ENZYME SUBSTRATE

Provided is a ultra-high-sensitivity assay in which the assay can be made on a commonly used assay apparatus such as an absorptiometer and a plate reader or with naked eyes. The high-sensitivity assay in which the assay can be made on a commonly used assay apparatus or with naked eyes can be provided by combining an enzyme cycling method using thio-NAD(P) as a coenzyme, a labeling enzyme and a substrate for the labeling enzyme optimally, and by amplifying thio-NAD(P)H, which is a signaling substance, exponentially and then quantifying the thio-NAD(P)H colorimetrically. 3. The method according to claim 1 , wherein:the enzyme is alkaline phosphatase, and{'sup': 1', '2, 'sub': 2-6', '1-6, 'X represents a phosphate group, Yrepresents hydrogen, and Yrepresents a Calkoxy group, or a Calkyl group in the androsterone derivative represented by the formula (1).'}4. The method according to claim 2 , wherein:the enzyme is alkaline phosphatase, and{'sup': 1', '2, 'sub': 2-6', '1-6, 'X represents a phosphate group, Yrepresents hydrogen, and Yrepresents a Calkoxy group, or a Calkyl group in the androsterone derivative represented by the formula (1).'}5. The method according to claim 1 , wherein:the enzyme is glucosidase, galactosidase, fructosidase or mannosidase, and{'sup': 1', '2, 'sub': 2-6', '1-6, 'X represents a sugar moiety, the sugar moiety represents one selected from the group consisting of glucose, galactose, fructose and mannose, and Yrepresents hydrogen, and Yrepresents a Calkoxy group, or a Calkyl group in the androsterone derivative represented by the formula (1).'}6. The method according to claim 2 , wherein:the enzyme is glucosidase, galactosidase, fructosidase or mannosidase, and{'sup': 1', '2, 'sub': 2-6', '1-6, 'X represents a sugar moiety, the sugar moiety represents one selected from the group consisting of glucose, galactose, fructose and mannose, and Yrepresents hydrogen, and Yrepresents a Calkoxy group, or a Calkyl group in the androsterone derivative ...

IMMOBILIZED PROTEIN SYSTEM FOR RAPID AND ENHANCED MULTIPLEXED DIAGNOSTICS

The disclosure relates to methods of detecting a neural injury biomarker in a biological sample. The method includes subjecting a biological sample to an assay disclosed that produces a measurable signal and detecting the measurable signal. The presence or absence of the measurable signal indicates the presence or absence of the neural injury biomarker in the sample, and thereby diagnosing a subject as having a neural injury. The disclosure further relates to methods of determining the state of a subject's neural injury. Further disclosed are systems and devices useful in carrying out the methods disclosed. 1. A method for detecting the presence or absence of a neural injury biomarker in a biological sample , the method comprising:providing a biological sample; [{'sup': +', '+, '(a) an assay to detect S100β, wherein the assay comprises (i) providing aldolase, glyceraldehyde 3-phosphate dehydrogenase (“GAPDH”), and a signal-transducing molecule, each immobilized on one or more supports, (ii) providing fructose 1,6-bisphosphate and NAD, and (iii) contacting the biological sample with the one or more supports, NAD, and fructose 1,6-bisphosphate under conditions effective to cause a cascading biological reaction leading to production of a measurable signal by the signal-transducing molecule if S100β is present in the biological sample;'}, '(b) an assay to detect S100β, wherein the assay comprises (i) providing phosphoglyceromutase (“PGM”), enolase, pyruvate kinase, and a signal-transducing molecule, each immobilized on one or more supports, (ii) providing 3-phosphoglycerate and (iii) contacting the biological sample with the one or more supports and 3-phosphoglycerate under conditions effective to cause a cascading biological reaction leading to production of a measurable signal by the signal-transducing molecule if S100β is present in the biological sample;', {'sub': 2', '2, '(c) an assay to detect glial fibrillary acidic protein (“GFAP”), wherein the assay comprises ( ...

Enzymatically responsive magnetic particles and their use

The invention relates to an enzymatically responsive product that includes an amino acid residue conjugated to a magnetic particle, wherein the amino acid residue is phosphorylated or sulfated or comprises an ester-moiety linked via peptide bond. Compositions containing the enzymatically responsive product, and the use thereof for separating distinct types of mammalian cells (e.g., cancer cells from normal cells), for treating a cancerous condition, and imaging cancer cells are also disclosed.

Nanoscale probe structure and application thereof

A nanoscale probe structure, including: a first probe having a tip top end and a second probe having a planar top end, wherein a metallic layer coats the on the tip top end, an insulating layer coats around the tip top end of the first probe; and a metallic layer coats on the planar top end, an insulating layer coats around the planar top end of the second probe. The structure of present invention can applied in atomic force microscopy to measure the electricity physiology signal inside and outside the cell membrane, which can limit the measure region to specific little area for the measure of electricity physiology signal and effectively decrease the miscellaneous noise disturbance from other region.

METHODS FOR ASSESSING TOXICITY

A method for characterizing toxicity of toxic pollutants and a method for characterizing comprehensive toxicity of water bodies. The methods include constructing a reporter gene cell line expressing CHOP gene associated with endoplasmic reticulum stress. 1. A method for constructing a reporter gene cell line expressing CHOP gene associated with endoplasmic reticulum stress , the method comprising:using the CHOP gene associated with endoplasmic reticulum stress as a specific inducing gene to construct a CHOP promoter;ligating the CHOP promoter with a SEAP gene to construct a lentivirus CHOP-SEAP plasmid vector; andtransfecting the CHOP-SEAP plasmid into Hela cells to construct the reporter gene cell line. Pursuant to 35 U.S.C. § 119 and the Paris Convention Treaty, this application claims foreign priority to Chinese Patent Application No. 201710616052.5 filed Jul. 25, 2017, the contents of which are incorporated herein by reference. Inquiries from the public to applicants or assignees concerning this document or the related applications should be directed to: Matthias Scholl P. C., Attn.: Dr. Matthias Scholl Esq., 245 First Street, 18th Floor, Cambridge, Mass. 02142.The present disclosure relates to the field of detection of cytotoxicity caused by contaminants, and more particularly to the use of a reporter gene cell line based on endoplasmic reticulum stress in characterizing toxicity of pollutants.Reporter gene assay is widely used in toxicity evaluation because of its sensitivity, rapidity, and reproducibility. Researches have shown that the reporter gene method can significantly lower the toxicity detection limit, improve the sensitivity, and shorten the response time. The current research based on reporter gene is mainly to monitor the specific target substances, such as DNA damaging substances, endocrine disruptors, dioxins, and other substances through the specific combination of inducible gene and reporter gene. However, different pollutants have different ...

MENAINV AND CANCER INVASION AND METASTASIS

Methods and compositions are provided for diagnosing or inhibiting invasion or metastasis of a cancer in a subject based on Mena. 1. A method of identifying an agent as an inhibitor of sarcoma or carcinoma cell invasion or as an inhibitor of metastasis of a carcinoma or sarcoma , comprising contacting a preparation comprising a growth factor receptor and a protein-tyrosine phosphatase 1b (PTP1b) in the presence of an amount of Menaand an amount of Mena and quantifying the association of the growth factor receptor and PTP1b in the presence of the agent and in the absence of the agent , wherein an agent that increases the association of growth factor receptor and PTP1b in the presence of the agent as compared to in the absence of the agent is identified as an inhibitor of sarcoma or carcinoma cell invasion or of metastasis , and an agent that does not increase , or decreases , the association of growth factor receptor and PTP1b in the presence of the agent as compared to in the absence of the agent is not identified as an inhibitor of sarcoma or carcinoma cell invasion or metastasis.2. A method of identifying an agent as a sensitizer of a growth factor receptor-expressing carcinoma cell to a corresponding growth factor or as a dysregulator of a PTP1b-binding growth factor receptor , comprising contacting a preparation comprising a growth factor receptor and a protein-tyrosine phosphatase 1b (PTP1b) in the presence of an amount of Menaand an amount of Mena and quantifying the association of the growth factor receptor and PTP1b in the presence of the agent and in the absence of the agent , wherein an agent that decreases the association of growth factor receptor and PTP1b as compared to the association in the absence of the agent is identified as a sensitizer of a growth factor receptor-expressing carcinoma cell to the corresponding growth factor or as a dysregulator , and an agent that does not change or that increases the association of growth factor receptor and ...

BIOMARKERS OF THERAPEUTIC RESPONSIVENESS

The present invention relates to methods of diagnosing breast cancer in a patient, as well as methods of monitoring the progression of breast cancer and/or methods of monitoring a treatment protocol of a therapeutic agent or a therapeutic regimen, The invention also relates to assay methods used in connection with the diagnostic methods described herein. 122-. (canceled)23. A method of administering a treatment regimen to a patient in need thereof for treating breast cancer , comprising:(a) measuring in a first test sample from a patient before said treatment regimen is initiated, baseline levels of a plurality of biomarkers comprising phosphorylated isoforms of Akt, MEK, mTOR, and GSK3beta,(b) measuring in an interim test sample from said patient during said treatment regimen for breast cancer interim levels of said plurality of biomarkers in said interim test sample,(c) comparing said interim levels to said baseline levels of said plurality of biomarkers,(d) evaluating from said comparing step (c) whether said patient is responsive to said treatment regimen, wherein if said interim levels of phosphorylated isoforms of Akt, MEK, mTOR, and GSK3beta are decreased as compared to said baseline levels, then the patient is responsive to said treatment regimen, and wherein if said interim levels of phosphorylated isoforms of Akt, MEK, mTOR, and GSK3beta are unchanged as compared to said baseline levels, then the patient is not responding to said treatment.24. The method of claim 23 , wherein each measuring step comprises conducting a multiplexed assay measurement of a plurality of said biomarkers in said test sample claim 23 , wherein said multiplexed assay measurement is conducted using one reaction volume comprising said test sample.25. The method of claim 23 , further comprising determining from said interim levels of said plurality of biomarkers the disease progression of breast cancer.26. The method of claim 23 , wherein each measuring step measures said level using ...

Patch-Sized Apparatus And Method For Use With An Apparel To Detect Seminal Fluid

The present invention is a patch-sized apparatus for use with an apparel to detect seminal fluid that includes disposable housing. The disposable housing is removably attached to the apparel. The disposable portion incorporates therein a fluid channel comprising a chemical solution to detect seminal fluid present on the surface of the apparel. The chemical solution includes but not limited to 1-Naphthylphosphate (disodium salt); and prostate-specific antigen (PSA)(A67-B/E3).

Screening Broths Comprising Esculatin Compounds for the Detection of Specific Microorganisms

The present invention relates to the use of an esculatin compound in a method for detecting the presence or absence of an enzymatic reaction in a sample containing one or more enzymes and one or more first chromogenic indicator of one or more enzymatic reactions, wherein the esculatin compound is a compound of formula 2. The method according to claim 1 , wherein the enzymatically cleavable group is an α or β linked sugar residue.3. The method according to claim 1 , wherein the enzymatically cleavable group is a phosphate group having the formula POW.4. The method according to claim 1 , wherein Rand Rtogether with the carbon atoms to which they are attached form a cyclohexene ring.7. The method according to claim 1 , wherein the one or more first chromogenic indicator of the enzymatic activity is a pH indicator claim 1 , a visible color indicator claim 1 , or a fluorescent indicator.8. The method according to claim 7 , wherein the one or more first chromogenic indicator of the enzymatic activity is 5-bromo-4-chloroindoxyl phosphate.9. The method according to claim 7 , wherein the one or more first chromogenic indicator of the enzymatic activity is 4-methylumbelliferyl-β-D glucopyranoside.10. The method according to claim 7 , wherein the one or more first chromogenic indicator of the enzymatic activity is 4-methylumbelliferyl-β-D glucuronide.11. The method according to claim 1 , wherein the dark precipitate masks the fluorescence or color of the one or more first chromogenic indicator.12. The method according to claim 1 , wherein the presence or absence of the dark precipitate is a secondary or confirmatory test after the one or more first chromogenic indicator.13staphylococcus aureus, listeria, salmonella, clostridium, streptococcus, klebsiella, enterobacter, escherichia, citrobacter, proteus, bacillus, pseudomonas, lactobacillus,. The method according to claim 1 , wherein the one or more microorganisms are bacterial microorganisms and are selected from and coliforms ...

DEGRADABLE CARBON NANOTUBE-CONTAINING BIOSENSORS AND METHODS FOR TARGET CLINICAL MARKER DETECTION

The invention relates to carbon nanotube-containing composites as biosensors to detect the presence of target clinical markers, methods of their preparation and uses in the medical field. The invention is particularly suitable for the detection in patient biological specimens of bone markers and tissue markers. The biosensors of the invention include carbon nanotubes deposited on a substrate, gold nanoparticles deposited on the carbon nanotubes and, binder material and biomolecule deposited on the gold-coated carbon nanotubes. The biomolecule is selected to interact with the target clinical markers. The biosensor can be used as an in-situ or an ex-situ device to detect and measure the presence of the target clinical markers.

Senescent Cell Biomarkers

The invention relates to senescent cell biomarkers and the uses thereof. The invention also extends to methods and kits for detecting senescence, and drug conjugates and pharmaceutical compositions for killing senescent cells. 1. A method of detecting a senescent cell in a sample , the method comprising detecting the expression , in the sample , of at least one senescent cell biomarker selected from DEP-1 , NTAL , EBP50 , STX4 , VAMP3 , ARMCX-3 , LANCL1 , B2MG , PLD3 and VPS26A , or a variant or fragment thereof , wherein an increased level of expression of the at least one biomarker or a variant or fragment thereof relative to the level of expression detected in a reference sample is an indication of a senescent cell present in the sample.2. A method according to claim 1 , wherein the at least one senescent cell biomarker comprises an amino acid sequence substantially as set out in any one of SEQ ID Nos. 1 to 19 claim 1 , or a variant or fragment thereof.3. A method according to claim 1 , wherein the at least one senescent cell biomarker is DEP-1 claim 1 , NTAL claim 1 , EBP50 claim 1 , STX4 claim 1 , VAMP-3 claim 1 , PLD3 or ARMCX-3 claim 1 , or a variant or fragment thereof.4. (canceled)5. A method according to claim 1 , wherein one or more of DEP-1 claim 1 , NTAL claim 1 , B2MG claim 1 , ARMCX-3 claim 1 , PLD3 and LANCL1 claim 1 , or a variant or fragment thereof claim 1 , is used as an extracellular biomarker.6. (canceled)7. (canceled)8. The method according to claim 1 , wherein the method comprises detecting two claim 1 , three claim 1 , four or more biomarkers claim 1 , or variants or fragments thereof claim 1 , in the sample.9. The method according to claim 1 , wherein the sample is a bodily sample taken from a test subject claim 1 , and wherein the test subject is an experimental animal or a human.10. (canceled)11. (canceled)12. The method according to claim 1 , wherein sample is an ex vivo sample or an in vitro sample.13. A senescent cell detection kit for ...

Senescent Cell Biomarkers

The invention relates to senescent cell biomarkers and the uses thereof. The invention also extends to methods and kits for detecting senescence, and drug conjugates and pharmaceutical compositions for killing senescent cells. 16-. (canceled)7. A method of detecting a senescent cell in a sample , the method comprises detecting the expression , in the sample , of at least one senescent cell biomarker selected from DEP-1 , NTAL , EBP50 , STX4 , VAMP3 , ARMCX-3 , LANCL1 , B2MG , PLD3 and VPS26A , or a variant or fragment thereof , wherein an increased level of expression of the at least one biomarker or a variant or fragment thereof relative to the level of expression detected in a reference sample is an indication of a senescent cell present in the sample.8. The method according to claim 7 , wherein the method comprises detecting two claim 7 , three claim 7 , four or more biomarkers claim 7 , or variants or fragments thereof claim 7 , in the sample.9. The method according to claim 7 , wherein the sample is a bodily sample taken from a test subject.10. The method according to claim 7 , wherein the sample comprises blood claim 7 , plasma claim 7 , serum claim 7 , spinal fluid claim 7 , urine claim 7 , sweat claim 7 , saliva claim 7 , tears claim 7 , breast aspirate claim 7 , prostate fluid claim 7 , seminal fluid claim 7 , vaginal fluid claim 7 , stool claim 7 , cervical scraping claim 7 , cytes claim 7 , amniotic fluid claim 7 , intraocular fluid claim 7 , mucous claim 7 , moisture in breath claim 7 , animal tissue claim 7 , cell lysates claim 7 , tumour tissue claim 7 , hair claim 7 , skin claim 7 , buccal scrapings claim 7 , nails claim 7 , bone marrow claim 7 , cartilage claim 7 , prions claim 7 , bone powder claim 7 , ear wax claim 7 , or combinations thereof.11. The method according to claim 9 , wherein the test subject is an experimental animal or a human.12. The method according to claim 7 , wherein sample is an ex vivo sample or an in vitro sample.13. A ...

DIAGNOSIS AND TREATMENT OF INCIPIENT DIABETES

A method is described for predicting incipient diabetes, metabolic disorders or the metabolic syndrome by developing a personal temporal Phosphatase profile, which is generated by measuring phosphatase concentration in stool at a single time-point or multiple time-points. The phosphatase profile further can be used for diagnosing and determining prognosis of other incipient or overt diseases, such as the metabolic syndrome, coronary heart disease, nonalcoholic fatty liver disease, cancers, other chronic or acute diseases or infectious diseases. Also described is a specific dose of phosphatase for therapeutic use in incipient diabetes and other incipient or overt diseases. 1. A method for developing a temporal profile of stool phosphatase comprising measuring concentration of phosphatase in a stool sample from a subject , wherein said stool sample is collected at a single time point or at multiple time points.2. The method of claim 1 , wherein said multiple time points comprise daily claim 1 , weekly claim 1 , monthly or yearly.3. The method of claim 2 , wherein said subject is a human claim 2 , cattle claim 2 , pig claim 2 , sheep claim 2 , goat claim 2 , cow claim 2 , horse claim 2 , dog claim 2 , cat claim 2 , monkey claim 2 , rabbit claim 2 , rat claim 2 , mouse claim 2 , chicken claim 2 , or turkey.4. The method of claim 3 , wherein said measuring comprises:providing a substrate for phosphatase in a solid or liquid form;contacting said substrate for phosphatase with a stool sample;waiting a specified period of time to allow a color to develop or waiting a specified period of time to allow an enzymatic reaction to occur;optionally, counting pixels of photographs;comparing the developed color with photographs of standards or comparing the number of pixels to standards; andquantifying the concentration of phosphatase in said stool sample or quantifying the concentration of phosphatase in said stool sample using a spectophotometer, a biochemistry analyzer, a high- ...

ASSAY FOR SHIP1 EXPRESSION, ACTIVITY AND SEQUENCE ALTERATIONS AS A PREDICTOR OF INFLAMMATORY BOWEL DISEASE RISK

The present disclosure is directed to detecting colon disorders by measuring the expression of SHIP1 in a sample of PBMCs. One method includes the following steps, obtaining a sample including peripheral blood mononuclear cells (PBMCs) from a subject and determining whether SHIP1 is underexpressed in the PBMCs or lacks normal enzymatic activity. The present disclosure is also directed to a method of determining the expression of SHIP1 protein expression and SHIP1 enzyme activity in PBMCs. This method includes the following steps, obtaining a sample comprising PBMCs from a subject and determining the amount of SHIP1 in the PBMCs. 1. A method of detecting a colon disorder in a subject comprising:obtaining a sample comprising peripheral blood mononuclear cells (PBMCs) of the subject; anddetermining levels of expression or enzymatic activity of SH2-containing inositol-5-phosphatase 1 (SHIP1) in the PBMCs, wherein underexpression or lack of a normal enzymatic activity is indicative of the colon disorder.2. The method of claim 1 , wherein underexpression of SHIP1 is indicative of the colon disorder.3. The method of claim 1 , wherein the subject is a human.4. The method of claim 1 , wherein the colon disorder is inflammatory bowel disease.5. The method of claim 4 , wherein the colon disorder is Crohn's Disease.6. The method of claim 1 , wherein the sample is a whole blood sample.7. A method of determining the expression of SH2-containing inositol-5-phosphatase 1 (SHIP1) protein expression and enzyme activity in peripheral blood mononuclear cells (PBMCs) claim 1 , comprising:obtaining a sample comprising PBMCs from a subject;determining the amount of SHIP-1 in the PBMCs; anddetermining the amount of enzyme activity in the PBMCs.8. A method of detecting a colon disorder in a subject comprising:obtaining a sample containing the gene phosphatidylinositol-3,4,5-trisphosphate 5-phosphatase 1(INPP5D) of the subject;determining whether the INPP5D includes a Single Nucleotide ...

Methods for the Treatment of Solid Tumor Cancers Using Illudins and Biomarkers

Methods for determining the likelihood that a subject suffering from a solid tumor cancer will benefit from treatment with an illudin are disclosed herein. Further, there are also methods for treatment based on such determination. In several embodiments, markers prostaglandin reductase 1 (PTGR1), Protein Tyrosine Phosphatase Non-Receptor Type 14 (PTPN14), Aspartate Beta-Hydroxylase (ASPH) together with one or more genes or alone may be used to enhance or guide treatment with an illudin. In certain embodiments, the protein or gene may be expressed or methylated. 2. The method of claim 1 , wherein the solid tumor cancer is prostate cancer claim 1 , ovarian cancer claim 1 , kidney cancer claim 1 , and thyroid cancer.3. The method of claim 1 , wherein the solid tumor cancer is colorectal cancer claim 1 , pancreatic cancer claim 1 , primary liver cancers claim 1 , kidney cancer claim 1 , ovarian cancer claim 1 , uterine cancer claim 1 , or breast cancer.4. The method of claim 1 , wherein the ASPH is methylated.6. The method of claim 5 , further comprising the step of measuring the level of expression of PTGR1 of a combination thereof in the biological sample.7. The method of claim 5 , further comprising the step of measuring the level of expression of PTPN14 in the biological sample.8. The method of claim 5 , further comprising e step of measuring the level of expression of ASPH in the biological sample.9. The method of claim 5 , wherein the cancer is newly diagnosed claim 5 , relapsed claim 5 , or refractory.10. The method of claim 5 , further comprising modifying a targeted drug therapy based on expression of a gene.11. The method of claim 9 , wherein ASPH is methylated.12. The method of claim 1 , wherein the cancer is colorectal cancer claim 1 , pancreatic cancer claim 1 , primary liver cancers claim 1 , kidney cancer claim 1 , ovarian cancer claim 1 , uterine cancer claim 1 , lung cancer claim 1 , breast cancer claim 1 , prostate cancer claim 1 , sarcomas claim 1 , ...

METHODS FOR DETECTING ORGANISMS AND ENZYMATIC REACTIONS USING RAMAN SPECTROSCOPY AND AROMATIC COMPOUNDS COMPRISING PHOSPHATE

The present disclosure provides systems for the rapid and sensitive detection of organisms and molecules in samples. Reactants that produce Raman-active products are used in combination with Raman light scattering. Such compounds may comprise phosphates permitting the detection of phosphatases. The present disclosure can also be used to measure enzyme-kinetics. 1. A method for detecting the activity of at least one enzyme in a sample comprising:a) preparing a mixture comprising the sample and at least one aromatic compound comprising at least one phosphate group; 'i) optionally adding an oxidizing agent; and', 'b) incubating the mixture in the presence of a base to form at least one Raman-active product;'}c) detecting the at least one Raman-active product with Raman spectroscopy,wherein the method does not comprise Surface Enhanced Resonance Raman Scattering.7. The method of claim 1 , wherein the at least one enzyme comprises a phosphatase.8. The method of claim 7 , wherein the phosphatase is alkaline phosphatase.9. The method of claim 8 , wherein the alkaline phosphatase is conjugated to an antibody.10. The method of claim 1 , wherein the at least one aromatic compound comprises 4-amino-1-phenyl-1-phosphate.11. The method of claim 1 , wherein the at least one aromatic compound comprises 4-hydroxy-1-naphthyl-1-phosphate.12. The method of claim 1 , wherein the at least one aromatic compound comprises 4-amino-1-naphthyl-1-phosphate.13. The method of claim 1 , wherein the at least one aromatic compound comprises hydroquinone diphosphate; and wherein the method comprises adding an oxidizing agent.14. The method of claim 1 , wherein the base is sodium hydroxide.15. The method of claim 1 , wherein the oxidizing agent is sodium metaperiodate.16. The method of claim 1 , wherein the Raman spectroscopy is resonant Raman spectroscopy.17. A method for detecting at least one target in a sample comprising:a) preparing a mixture comprising the at least one target;b) incubating the ...

ASSAY FOR SCREENING MODULATORS OF HOLOPHOSPHATASE ACTIVITY

This invention relates to assays for screening test compounds for their ability to modulate holophosphatase activity. 2. A method according to wherein the modulation of holophosphatase activity is inhibition of holophosphatase activity.3. A method according to wherein the modulation of holophosphatase activity is activation of holophosphatase activity.4. A method according to any preceding claim wherein the holophosphatase activity that is modulated by the test compound is selective holophosphatase activity.5. A method according to any preceding claim wherein the holophosphatase is purified.6. A method according to any preceding claim wherein the holophosphatase claim 1 , the catalytic subunit claim 1 , the at least one regulatory subunit and/or the phosphorylated substrate is a recombinant protein.7. A method according to wherein the holophosphatase is synthesised by expressing the catalytic subunit and the at least one regulatory subunit in a cell system so as to generate a functional and selective reconstituted form.8. A method according to any preceding claim wherein the regulatory subunit is a truncated fragment of a naturally occurring regulatory subunit.9. A method according to any preceding claim wherein the catalytic subunit comprises a Ser/Thr phosphoprotein phosphatase (PPP).10. A method according to any preceding claim wherein the catalytic subunit comprises a Ser/Thr protein phosphatase 1 subunit (PP1).11. A method according to any preceding claim wherein the catalytic subunit comprises a PP1c subunit.12. A method according to any preceding claim wherein the regulatory subunit is selected from R15A and R15B claim 6 , or fragments thereof.13. A method according to wherein the fragment of a regulatory subunit is selected from R15Aand R15B.14. A method according to wherein the fragment of a regulatory subunit is selected from R15A(R15A) claim 12 , R15A(R15A) claim 12 , R15B(R15B) or R15B(R15B).15. A method according to any of to wherein the regulatory ...

Fusion peptides or proteins, their use, and systems and kits based thereupon, for the separation and/or detection of plastics, particularly of microplastics

The present invention pertains to a novel fusion protein and/or fusion peptide, preferably for use in the separation from and/or detection in an environment of one or more target polymers or plastics, e.g., one or more target polymer fragments and/M or particles or target plastic fragments and/or particles, preferably wherein the one or more target polymer particles or target plastic particles are microplastics; a method of preparing such novel fusion protein and/or fusion peptide, a system and kit comprising the novel fusion protein and/or fusion peptide and a (polymer or non-polymer) carrier or carrier system, a use of the novel fusion protein and/or fusion peptide or of a system and kit as mentioned in the separation from and/or detection in an environment of one or more target polymers or plastics, e.g., one or more target polymer fragments and/or particles or target plastic fragments and/or particles, preferably wherein the one or more target polymer particles or target plastic particles are microplastics; a method of separation of one or more target polymers or plastics from an environment, e.g., one or more target polymer fragments and/or particles or target plastic fragments and/or particles, and a method of detection of one or more target polymers or plastics in an environment, e.g., one or more target polymer fragments and/or particles or target plastic fragments and/or particles, preferably wherein the one or more target polymer particles or target plastic particles are microplastics.

METHODS TO ASSAY KINASE ACTIVITY

Methods and kits for enzymes involved in post-translational modifications are provided. The methods employ elemental analysis, including ICP-MS. The methods allow for the convenient and accurate analysis of post-translation modifications of substrates by enzymes involved in post-translational modifications, including kinase and phosphatase enyzmes 1. A method for a kinase assay , comprising:a) Incubating ATP, at least one kinase, and a free non-phosphorylated substrate labeled with an element tag, with a support having attached thereto metal ion coordination complexes under conditions to enable the kinase to phosphorylate the substrate;b) Separating free non-phosphorylated substrate from bound phosphorylated substrate labeled with an element tag to the support;c) Eluting the element tag associated with the resultant phosphorylated substrate into a solution; andd) Performing solution elemental analysis of said solution.2. The method of where in step (a) a multitude of free non-phosphorylated substrates each labeled with a unique element tag are incubated.3. The method of wherein the metal ion coordination complex attached to the support is a titanium oxide bead.4. The method of where in step (a) the conditions to enable the kinase to phosphorylate the substrate include a kinase reaction buffer.5. The method of where in step (a) antagonists or agonists of kinase are incubated.6. The method of wherein the kinase is delivered in the form of a cell lysate.7. A method for a phosphatase assay claim 1 , comprising:a) Incubating free phosphorylated substrate labeled with an element tag with a support having attached thereto metal ion coordination complexes;b) Separating free phosphorylated substrate from bound phosphorylated substrate labeled with an element tag attached to the metal ion coordination complexes attached to the support;c) Incubating ADP and at least one phosphatase with the bound phosphorylated substrate labeled with an element tag attached to the metal ion ...

ISOLATION OF ADULT MULTIPOTENTIAL CELLS BY TISSUE NON-SPECIFIC ALKALINE PHOSPHATASE

The present invention relates to the use of tissue non-specific alkaline phosphatase (TNAP) as a marker for identifying and/or isolating adult multipotential cells. The present invention also relates to cell populations enriched by methods of the present invention and therapeutic uses of these cells. 1. Use of TNAP as a marker for the identification and/or enrichment of adult multipotential cells.217-. (canceled)18. An enriched population of adult multipotential cells obtained by a method according to .19. An enriched population of TNAP+ adult multipotential cells.20. An expanded cell population obtained by culturing an enriched population of adult multipotential cells according to .2124-. (canceled)25. A composition comprising a population of enriched adult multipotential cells according to .2628-. (canceled)29. A method for generating or repairing tissue in a subject claim 18 , the method comprising administering to the subject an enriched or expanded cell population according to .30. A method for generating or repairing tissue in a subject claim 25 , the method comprising administering to the subject a composition according to .31. A method according to wherein the tissue is bone claim 29 , cardiac or cartilage tissue.3240-. (canceled)41. A STRO-3 hybridoma cell line deposited with ATCC on 19 Dec. 2005 under the provisions of the Budapest Treaty under deposit accession number PTA-7282.42. A STRO-3 antibody produced by the hybridoma cell line deposited with ATCC on 19 Dec. 2005 under the provisions of the Budapest Treaty under deposit accession number PTA-7282.43. An isolated antibody which binds to the same epitope on multipotential cells as the STRO-3 antibody produced by the hybridoma cell line deposited with ATCC on 19 Dec. 2005 under the provisions of the Budapest Treaty under deposit accession number PTA-7282.44. A composition comprising an antibody according to or .45. (canceled)46. A method for generating or repairing tissue in a subject claim 19 , the ...

MODULATORS OF PROTEIN TYROSINE PHOSPHATASE AND USES THEREOF

Composition and methods are provided for the specific manipulation of protein tyrosine phosphatase (PTP) activity, including without limitation manipulation of protein tyrosine phosphatase receptor type gamma (PTPRG). The modulation of PTP activity can be performed in vitro or in vivo, and is useful for therapeutic and research purposes. In some embodiments, an effective dose of a PTP modulator is provided to an individual for preventing or treating disease involving dysregulated tyrosine kinase activity and/or signaling mechanisms involving tyrosine phosphorylation and/or tyrosine kinase activity. In other embodiments, a PTP modulator is utilized in the analysis and screening of phosphatase pathways in a cell. 1. A method of increasing protein tyrosine phosphatase receptor type gamma (PTPRG) activity in a targeted cell population , the method comprising:contacting cells in the targeted population with an effective dose of a protein tyrosine phosphatase (PTP) activating agent selected from a PTP wedge domain (WD) polypeptide; a PTP intracellular domain (ICD) polypeptide; a PTP intracellular phosphatase domain (IPD) peptide; and a PTP soluble extracellular domain polypeptide (PTPx); or fusion, variant, or mimetic thereof;wherein PTPRG activity in the cell is increased.2. The method of claim 1 , wherein the effective dose increases PTPRG activity to result in altered tyrosine phosphorylation of one or more target proteins.3. The method of claim 1 , wherein the PTP activating agent is a human polypeptide or a fusion polypeptide thereof claim 1 , optionally fused to a permeant domain.4. (canceled)5. The method of claim 1 , wherein endogenous PTPRG protein in the cell is activated.6. The method of claim 1 , wherein the PTP activating agent is a human PTPRG ICD or IPD polypeptide or fusion polypeptide thereof.7. The method of claim 6 , wherein the ICD or IPD provides phosphatase activity in the absence of endogenous PTPRG.8. The method of claim 1 , wherein the targeted ...

Biomarkers of therapeutic responsiveness

The present invention relates to methods of diagnosing breast cancer in a patient, as well as methods of monitoring the progression of breast cancer and/or methods of monitoring a treatment protocol of a therapeutic agent or a therapeutic regimen. The invention also relates to assay methods used in connection with the diagnostic methods described herein.

METHOD FOR DETECTING FLUORESCENCE OR ABSORBANCE, METHOD FOR SUPPRESSING BACKGROUND, METHOD FOR MEASURING ADP, METHOD FOR MEASURING ACTIVITY OF ADP-SYNTHESIZING ENZYME, AND METHOD FOR MEASURING ACTIVITY OF GLUCOSYLTRANSFERASE

In a method for detecting fluorescence or absorbance of the present invention, a diaphorase causes reduction from resazurin to resorufin in the presence of an SH reagent and NADH or NADPH, and the resulting fluorescence intensity or absorbance is measured. A method for measuring ADP of the present invention includes a 2-1 process in which glucose is reacted with ADP and an ADP-dependent hexokinase, a 2-2 process in which the glucose-6-phosphate obtained in the 2-1 process is reacted with NAD or NADP and glucose-6-phosphate dehydrogenase, and a 2-3 process in which resazurin is reacted with the NADH or NADPH obtained in the 2-2 process and a diaphorase in the presence of an SH reagent, and the resulting fluorescence intensity or absorbance is measured. 2. The method for detecting fluorescence or absorbance according to claim 1 , wherein the maleimide compound represented by General Formula [1] is N-ethylmaleimide claim 1 , maleimide or N-(2-sulfoethyl)maleimide.4. The method for suppressing background according to claim 3 , wherein the maleimide compound represented by General Formula [1] is N-ethylmaleimide claim 3 , maleimide claim 3 , or N-(2-sulfoethyl)maleimide.6. (canceled)7. The method for measuring ADP according to claim 5 , wherein the maleimide compound represented by General Formula [1] is N-ethylmaleimide claim 5 , maleimide or N-(2-sulfoethyl)maleimide.9. (canceled)10. The method for measuring activities of ADP-producing enzymes according to claim 8 , wherein the maleimide compound represented by General Formula [1] is N-ethylmaleimide claim 8 , maleimide or N-(2-sulfoethyl)maleimide.11. The method for measuring activities of ADP-producing enzymes according to claim 8 , wherein the ADP-producing enzyme is at least one type selected from the group including kinases claim 8 , ATPases claim 8 , nitrogenases claim 8 , tetrahydrofolate synthases claim 8 , acetyl-CoA carboxylase claim 8 , pyruvate carboxylase claim 8 , and glutathione synthase.13. (canceled)14 ...

METHODS FOR DIAGNOSIS, PROGNOSIS AND METHODS OF TREATMENT

This invention is directed to methods and compositions for diagnosis, prognosis and for determining methods of treatment. The physiological status of a cell present in a sample (e.g. clinical sample) can be used in diagnosis or prognosis of a condition (e.g. Chronic Lymphocytic Leukemia), in patient selection for therapy, to monitor treatment and to modify or optimize therapeutic regimens. 132.-. (canceled)33. A method of correlating and/or classifying an activation state of a CLL cell with a clinical outcome in an individual by:(i) subjecting the CLL cell from the individual to a modulator, where the CLL cell expresses a B-Cell receptor (BCR);(ii) determining the activation levels of a plurality of activatable elements; and(ii) identifying a pattern of the activation levels of the plurality of activatable elements to determine the presence or absence of an alteration in signaling proximal to BCR, where the presence of the alteration is indicative of a clinical outcome.34. The method of claim 33 , further comprising determining the level of one or more cell surface markers on the cell.35. The method of claim 34 , wherein the cell surface marker is selected from the group consisting of CD1 claim 34 , CD2 claim 34 , CD3 claim 34 , CD4 claim 34 , CD5 claim 34 , CD8 claim 34 , CD10 claim 34 , CD14 claim 34 , CD19 claim 34 , CD20 claim 34 , CD22 claim 34 , CD23 claim 34 , CD40 claim 34 , CD52 claim 34 , CD100 claim 34 , CD280 claim 34 , CD281 claim 34 , CD282 claim 34 , CD283 claim 34 , CD284 claim 34 , and CD289.36. The method of claim 35 , wherein the cell surface marker is selected from the group consisting of CD45 claim 35 , CD5 claim 35 , CD14 claim 35 , CD19 claim 35 , CD20 claim 35 , CD22 claim 35 , CD23 claim 35 , CD27 claim 35 , CD37 claim 35 , CD40 claim 35 , CD52 claim 35 , CD79 claim 35 , CD38 claim 35 , CD96 claim 35 , MEW Class I claim 35 , and MEW Class 2.37. The method of claim 34 , wherein the cell surface marker is selected from the group consisting of ...

Processes for increasing extraction of enzymes from animal feed and measuring activity of the same

Methods of increasing extraction of enzymes from animal feed and measuring enzyme activity are described herein.

METHODS FOR MEASURING ENZYME ACTIVITY USEFUL IN DETERMINING CELL VIABILITY IN NON-PURIFIED SAMPLES

An assay kit of reagents including a nucleic acid capable of acting as substrate for polymerase microorganism activity useful in a method of detecting polymerase activity as an indicator of the presence of a micro-organism in a sample are disclosed. The disclosed embodiments also relate to reagents for use in such methods, and to test kits comprising such reagents for carrying out the methods. 1. An assay kit comprised of reagents including a nucleic acid capable of acting as substrate for polymerase microorganism activity useful in a method of detecting polymerase activity as an indicator of the presence of a micro-organism in sample , comprising:(a) a polymerase specific substrate; and(b) differential cell lysis reagents to allow only viable micro-organism derived polymerase activity to modify the polymerase specific substrate.2. A method of detecting polymerase activity as an indicator of the presence of a micro-organism in a sample , comprising:(a) contacting the sample with a nucleic acid molecule which acts as a substrate for polymerase activity in the sample; and(b) determining, specifically, the presence of a nucleic acid molecule resulting from the action of the micro-organism polymerase on the substrate nucleic acid molecule, to thereby indicate the presence of the micro-organism, for screening normally sterile body fluids for the presence or absence of micro-organisms therein and to provide diagnostic patient management information.3. The method of claim 2 , wherein the determining step includes determining the presence and the amount of a nucleic acid molecule resulting from the action of the micro-organism polymerase on the substrate nucleic acid molecule.4. The method of claim 2 , wherein the determining step includes determining the amount of a nucleic acid molecule resulting from the action of the micro-organism polymerase on the substrate nucleic acid molecule. This application is a continuation-in-part of and claims the benefit of pending ...

Phosphate and tensin homolog (PTEN) for the detection of autoimmune diseases or conditions

A method for the detection of impaired responsiveness of CD4+ T-cells to regulatory T-cells (Treg), referred to as Treg resistance. The method includes measuring the expression levels of phosphatase and tension homolog (PTEN) in activated CD4+ T-cells. Furthermore, a screening method for the detection of an autoimmune disease or a condition, may comprise the steps of generating a functional gene expression profile by measuring the expression levels of phosphatase and tension homolog (PTEN) in Treg-resistant CD4+ T-cells from patients suffering of an autoimmune disease or condition, and comparing the obtained gene expression profile with the expression profile from Treg-sensitive CD4+ T-cells from healthy controls. PTEN can be utilized in a screening system for the detection of impaired responsiveness of CD4+ T-cells to Treg. 1. A method for the detection of impaired responsiveness of CD4+ T-cells to regulatory T-cells (Treg) , referred to as Treg resistance , by measuring the expression levels of phosphatase and tensin homolog (PTEN) in activated CD4+ T-cells.2. The method according to claim 1 , wherein the expression levels of PTEN are compared between activated CD4+ T-cells from patients and activated Treg-sensitive CD4+ and CD8+ T-cells from healthy donors claim 1 , wherein a downregulation of PTEN within activated CD4+ T-cells as compared to the activated Treg-sensitive CD4+ T-cells is indicative for Treg resistance.3. The method according to claim 1 , wherein an upregulation of PTEN is correlated with a responsiveness of activated CD4+ T-cells to Treg-mediated suppression.4. The method according to claim 1 , wherein the Treg resistance correlates with an accelerated IL-6 production after T cell receptor (TCR) stimulation claim 1 , enhanced phosphorylation of PKB/c-Akt and/or an increased IL-6 receptor (IL-6R) expression.5. The method according to claim 1 , wherein impaired responsiveness of CD4+ T-cells is restored by normalizing PTEN expression in activated ...

COMPARTMENTALISED SCREENING BY MICROFLUIDIC CONTROL

The invention describes a method for the identification of compounds which bind to a target component of a biochemical system or modulate the activity of the target, comprising the steps of: a) compartmentalising the compounds into microcapsules together with the target, such that only a subset of the repertoire is represented in multiple copies in any one microcapsule; and b) identifying the compound which binds to or modulates the activity of the target; wherein at least one step is performed under microfluidic control. The invention enables the screening of large repertoires of molecules which can serve as leads for drug development. 1115-. (canceled)116. A method for screening a repertoire of compounds for a compound having a desired activity , comprising the steps of:(a) providing an aqueous fluid comprising a repertoire of compounds;(b) compartmentalizing the repertoire of compounds into microcapsules by partitioning the aqueous fluid with an immiscible fluid as the aqueous fluid is flowing through a microfluidic channel, such that only one compound from the repertoire of compounds is represented in any one microcapsule, and further wherein each microcapsule comprises a target cell; and(c) determining an effect of one or more of the repertoire of compounds on the target cell by using a cell-based assay that produces a different reaction product in each microcapsule depending on an identity of the compound compartmentalized in the each microcapsule.117. The method of claim 116 , wherein the target cells are compartmentalized together with the repertoire of compounds in the microcapsules.118. The method of claim 116 , wherein the repertoire of compounds are attached to microbeads.119. The method of claim 118 , wherein the repertoire of compounds are attached to microbeads through one or more cleavable linkers.120. The method of claim 118 , wherein each microbead comprises a detectable tag.121. The method of claim 118 , further comprising releasing the compounds ...

COMBINATION PRODUCT FOR DETECTING TARGET MARKER

The present invention relates to a combination product for detecting a target marker simply and with high sensitivity. More specifically, the present invention relates to a combination product for detecting a target marker in a biological sample in combination with a target marker binding molecule which is capable of binding specifically to the target marker in the biological sample, the combination comprising, at least: 120-. (canceled)21. A combination product for detecting a target marker in a biological sample in combination with a target marker binding molecule which is capable of binding specifically to the target marker , the combination product comprising , at least:(a) a first binding agent comprising a first binding molecule which is capable of directly or indirectly binding specifically to the target marker binding molecule, and a labeling substance;(b) a linker molecule which is capable of binding specifically to the first binding agent; and(c) a second binding agent which is capable of binding specifically to the linker molecule, and comprises a second binding molecule and a labeling substance.22. The combination product according to claim 21 , wherein the linker molecule is capable of binding specifically to the labeling substance.23. The combination product according to claim 21 , wherein the linker molecule is an antibody or an antigen binding fragment thereof.24. The combination product according to claim 21 , wherein the labeling substance is at least one selected from a chemiluminescent label claim 21 , a metal particle claim 21 , a fluorescent label claim 21 , an enzyme label claim 21 , a coenzyme label claim 21 , a labeled antibody claim 21 , a dye claim 21 , a bioluminescent label claim 21 , a hapten and a polymer particle.25. The combination product according to claim 21 , wherein the first binding agent is a structure in which the first binding molecule and the labeling substance are connected with each other directly or indirectly via a ...

DEGRADABLE CARBON NANOTUBE-CONTAINING BIOSENSORS AND METHODS FOR TARGET CLINICAL MARKER DETECTION

The invention relates to carbon nanotube-containing composites as biosensors to detect the presence of target clinical markers, methods of their preparation and uses in the medical field. The invention is particularly suitable for the detection in patient biological specimens of bone markers and tissue markers. The biosensors of the invention include carbon nanotubes deposited on a substrate, gold nanoparticles deposited on the carbon nanotubes and, binder material and biomolecule deposited on the gold-coated carbon nanotubes. The biomolecule is selected to interact with the target clinical markers. The biosensor can be used as an in-situ or an ex-situ device to detect and measure the presence of the target clinical markers. 1. A degradable biosensor composite to detect a target clinical marker generated in a body of a patient , comprising:a substrate having a surface;a plurality of carbon nanotubes, having an other surface, deposited on the surface of the substrate;a plurality of gold nanoparticles electrodeposited on the outer surface of the plurality of carbon nanotubes;a binding material adsorbed on the plurality of gold nanoparticles; anda biotinylated biomolecule selected from the group consisting of antibody and aptamer, to bind with the binding material and interact with the target clinical marker selected from the group consisting of a bone marker and tissue marker.23-. (canceled)4. The biosensor of claim 1 , wherein the biotinylated biomolecule is aptamer.56-. (canceled)7. The biosensor composite of claim 1 , further comprising depositing or embedding the biosensor composite on a surface of a degradable implant device.8. The biosensor composite of claim 7 , wherein the implant device is a degradable scaffold.914-. (canceled)15. A method of preparing a degradable biosensor composite to detect a target clinical marker in a body of a patient claim 7 , comprising:providing a substrate having a surface;depositing on the surface of the substrate a plurality of ...

ISOLATION OF ADULT MULTIPOTENTIAL CELLS BY TISSUE NON-SPECIFIC ALKALINE PHOSPHATASE

The present invention relates to the use of tissue non-specific alkaline phosphatase (TNAP) as a marker for identifying and/or isolating adult multipotential cells. The present invention also relates to cell populations enriched by methods of the present invention and therapeutic uses of these cells. 145-. (canceled)46. An enriched population of TNAP adult multipotential cells , wherein at least 4% of the total cell population are TNAP , STRO-1 multipotential cells capable of being cultured in vitro to produce adipocytes , osteocytes and chondrocytes , wherein the TNAP marker is a TNAP marker which binds the STRO-3 antibody produced by the hybridoma cell line deposited under ATCC Accession Number PTA-7282.47. An enriched population according to claim 46 , wherein at least 10% of the total cell population are TRAP claim 46 , STRO-1 multipotential cells capable of being cultured in vitro to produce adipocytes claim 46 , osteocytes and chondrocytes.48. An enriched population according to claim 46 , wherein at least 20% of the total cell population are TRAP claim 46 , STRO-1 multipotential cells capable of being cultured in vitro to produce adipocytes claim 46 , osteocytes and chondrocytes.49. An enriched population according to claim 46 , wherein the STRO-1 cells are STRO-1.50. An enriched population according to claim 47 , wherein the STRO-1 cells are STRO-1.51. An enriched population according to claim 48 , wherein the STRO-1 cells are STRO-1.52. A method of generating a committed cell population of a specific tissue type claim 46 , the method comprising culturing a population of adult multipotential cells according to in the presence of one or more stimulatory factors; and subjecting said cultured population to conditions biasing differentiation of the adult multipotential cells to the specific tissue type.53. A method of generating a committed cell population of a specific tissue type claim 47 , the method comprising culturing a population of adult multipotential ...

MASS SPECTROMETRY METHOD FOR MEASURING THIAMINE IN BODY FLUID

Provided are methods for determining the amount of total thiamine in a body fluid sample using liquid chromatography and mass spectrometry. Total thiamine is converted to free thiamine by treatment with an acid phosphatase prior to thiamine separation and quantification. 1. A method for determining the amount of total thiamine in a plasma or serum sample , comprising:(i) performing an organic solvent extraction of a plasma or serum sample, wherein the result of the extraction is an organic solvent phase and an aqueous phase;(ii) purifying thiamine from the aqueous phase of by liquid chromatography;(iii) ionizing the purified thiamine by electrospray ionization (ESI) in positive ion mode;(iv) determining the amount of thiamine by mass spectrometry, wherein the amount of total thiamine in the sample is determined.2. The method of claim 1 , the method further comprising removing soluble protein from a plasma or serum sample;3. The method of claim 2 , wherein soluble protein is removed by treating said sample with an acid.4. The method of claim 1 , the method further comprising incubating the sample with an acid phosphatase claim 1 , for not longer than about 2 hours to convert phosphorylated thiamine to thiamine;5. The method of claim 4 , wherein the incubation is carried out at a pH of about 4.6±0.1.6. The method of claim 4 , wherein the incubation is carried out at a temperature of about 40° C.7. The method of claim 4 , wherein the incubation is carried out between about 1 and about 2 hours.8. The method of claim 1 , wherein the liquid chromatography comprises high performance liquid chromatography (HPLC).9. The method of claim 1 , wherein the ionizing comprises ionizing a parent ion having a mass/charge ratio of 265.00±1.0.10. The method of claim 9 , wherein the parent ion is fragmented into one or more daughter ions and wherein the amount of one or more of said daughter ions is determined.11. The method of claim 10 , wherein the one or more daughter ions comprises ...

COMPARTMENTALISED SCREENING BY MICROFLUIDIC CONTROL

The invention describes a method for the identification of compounds which bind to a target component of a biochemical system or modulate the activity of the target, comprising the steps of: a) compartmentalising the compounds into microcapsules together with the target, such that only a subset of the repertoire is represented in multiple copies in any one microcapsule; and b) identifying the compound which binds to or modulates the activity of the target; wherein at least one step is performed under microfluidic control. The invention enables the screening of large repertoires of molecules which can serve as leads for drug development. 1. A method for identifying a compound or compounds in a repertoire of compounds , which compound or compound(s) possess(es) a desired activity , comprising the steps of:a) compartmentalising the compounds into microcapsules, such that only a subset of the repertoire is represented in multiple copies in any one microcapsule; andb) identifying the compounds which possess the desired activity,wherein one or both of steps a) and b) is performed under microfluidic control.2. A method according to claim 1 , wherein step (a) comprises forming groups of microcapsules comprising individual compounds and mixing the groups of microcapsules to form an emulsified compound repertoire wherein a subset of the repertoire is represented in multiple copies in any one microcapsule.4. A method according to claim 1 , wherein the desired activity is selected from the group consisting of a binding activity and the modulation of the activity of a target.5. A method according to claim 4 , wherein the binding activity is binding to a target.6. A method according to claim 4 , wherein the modulated activity is a binding activity.7. A method according to claim 4 , wherein the modulated activity is a catalytic activity.8. A method according to claim 1 , wherein the compound reacts with a target to generate a reaction product.9. A method according to claim 3 , ...

ANIMAL FEED ENZYME EXTRACTION

An in-feed assay and buffers for extracting an enzyme from a feed pellet treated at high temperature is provided. Another embodiment of the invention is measuring the enzyme activity of the enzyme additive extracted from the animal feed, measuring the quantity of enzyme extracted from the animal feed, or measuring both. In one embodiment the enzyme is an animal feed additive, such as a phytase. 1. An aqueous composition for extraction of a polypeptide from a heat-treated solid , said aqueous composition comprising a bile-salt detergent , a denaturant , a base , and water.2. The composition of claim 1 , wherein said denaturant is a chaotropic agent.36.-. (canceled)7. The composition of claim 1 , wherein said base is a bicarbonate salt.810.-. (canceled)11. The composition of claim 1 , wherein said aqueous composition has a basic pH.1214.-. (canceled)15. The composition of claim 1 , wherein said bile-salt detergent comprises a steroid acid selected from the group consisting of taurocholic acid claim 1 , glycocholic acid claim 1 , cholic acid claim 1 , deoxycholic acid claim 1 , lithocholic acid claim 1 , chenodeoxycholic acid claim 1 , and any combination thereof.16. The composition of claim 1 , wherein said bile-salt detergent comprises a sodium cation.1720.-. (canceled)21. A method of extracting a polypeptide from a heat-treated feed pellet claim 1 , comprising:providing a heat-treated feed pellet comprising said polypeptide, said heat-treated pellet having been subjected to a heat treatment of at least 70° C.,contacting said heat-treated feed pellet with an aqueous solution,agitating said heat-treated feed pellet in contact with said aqueous solution to extract said polypeptide from said heat-treated feed pellet, andremoving said heat-treated feed pellet so as to obtain said extracted polypeptide in said aqueous solution.22. The method of claim 21 , wherein said aqueous solution comprises at least one detergent at a level of about or greater than a critical micelle ...

PHOSPHATASE OR KINASE ACTIVITY DETECTION COMPOSITION AND DETECTION METHOD

The present invention relates to a composition for detecting phosphatase or kinase activity and a method of detecting phosphatase or kinase activity. The kinase or phosphatase activity may be quantitatively measured in real time by using the composition of the present invention. 1. A composition for measuring kinase or phosphatase activity , the composition comprising (a) zinc ions (Zn) , (b) a zinc ion receptor comprising a chelating ligand , and (c) a kinase or phosphatase peptide substrate , and exhibiting a phosphorylation or dephosphorylation detection signal according to a change in a phosphorylation state of the peptide substrate.2. The composition of claim 1 , wherein the peptide substrate comprises any one selected from (a) a fluorescence signal-generating donor fluorophore and (b) a fluorophore acceptor that quenches a fluorescence signal by causing fluorescence resonance energy transfer (FRET) with the donor fluorophore claim 1 , and the zinc ion receptor comprises any other one not selected from a donor fluorophore and a fluorophore acceptor.3. The composition of claim 2 , wherein the peptide substrate is immobilized on a support.4. The composition of claim 3 , wherein the peptide substrate further comprises biotin claim 3 , and the support is NeutrAvidin agarose beads.5. The composition of claim 1 , wherein the peptide substrate comprises a polyhistidine claim 1 , and the zinc ion receptor comprising a chelating ligand is metal nanoparticles surface-modified with a chelating ligand.6. The composition of claim 5 , wherein the metal nanoparticles are any one selected from the group consisting of gold claim 5 , silver claim 5 , copper claim 5 , platinum claim 5 , palladium claim 5 , nickel claim 5 , and iron claim 5 , or a mixture of two or more thereof.7. The composition of claim 5 , wherein the metal nanoparticles have an average diameter of 2 nm to 50 nm.8. The composition of claim 5 , wherein the chelating ligand is any one or more selected from the ...

DIAGNOSTIC AND THERAPEUTIC METHODS AND COMPOSITIONS INVOLVING PTEN AND BREAST CANCER

Patients with ErbB2-overexpressing cancers can be given an ErbB2 targeting agent as a therapeutic regimen but not all patients are responsive. The present invention concerns the diagnostic, prognostic and therapeutic methods and compositions for evaluating potential efficacy of an ErbB2 targeting agent in ErbB2-overexpressing cancers by evaluating PTEN expression, which is predictive of responsiveness or resistance to ErbB2 targeting agents such as trastuzumab. Low PTEN expression is predictive of a patient who will respond poorly to trastuzumab. 1. A method for administering a treatment regimen comprising an effective amount of a PI3K inhibitor , said method comprising:(1) assaying a patient sample comprising at least one test cancer cell to evaluate PTEN expression in said at least one test cancer cell;(2) comparing the PTEN expression in (1) to a reference PTEN expression in reference cancer cells of a reference cohort of patients who are not candidates for said treatment regimen; and(3) administering said treatment regimen to a test patient for whom said at least one test cancer cell is evaluated in (1) to have PTEN expression that is lower than the reference PTEN expression in (2).2. The method of claim 1 , wherein said PTEN expression in (1) is no more than 50% said PTEN expression in (2).3. The method of claim 1 , wherein the at least one test cancer cell is selected from the group consisting of breast cancer claim 1 , lung cancer claim 1 , ovarian cancer claim 1 , brain cancer claim 1 , gastrointestinal tract cancer claim 1 , salivary duct cancer claim 1 , endometrial cancer claim 1 , prostate cancer claim 1 , head & neck cancer claim 1 , glioma claim 1 , pancreatic cancer claim 1 , hepatocellular cancer claim 1 , myeloma claim 1 , soft tissue sarcoma claim 1 , and non-small cell lung cancer.4. The method of claim 1 , wherein the at least one test cancer cell is breast cancer.5. The method of claim 1 , wherein PTEN expression is evaluated by sequencing PTEN ...

THERMOSTABLE HALOARCHAEAL INORGANIC PYROPHOSPHATASE

The invention pertains to a PPA from a microorganism belonging to the family Halobacteriaceae (HPPA), for example, a PPA from The HPPA provided by the invention is soluble, thermostable and active at high concentrations of salt and/or organic solvent. An embodiment of the invention provides a method of increasing the rate of a reaction by adding an HPPA to the reaction mixture, wherein the reaction produces PPi, for example, an enzymatic reaction, and wherein the reaction is carried out at moderately high temperature and/or low water activity. Further embodiments of the invention provide an assay to detect the PPi released during a reaction which produces PPi by adding an HPPA to convert the PPi in to Pi and measuring the resultant Pi. The invention further pertains to an assay to monitor a reaction which produces PPi in the presence or the absence of an HPPA. 143-. (canceled)44. An isolated inorganic pyrophosphatase from a microorganism belonging to family Halobacteriaceae (HPPA).45. The HPPA of claim 44 , wherein the HPPA is encoded by SEQ ID NO: 1 and has the sequence of SEQ ID NO: 2.46. The HPPA of claim 44 , wherein the HPPA has the sequence of SEQ ID NO: 3 to 108.47. An isolated nucleotide sequence encoding the HPPA of .48. A DNA construct comprising the nucleotide sequence of .49. A cell comprising the nucleotide sequence of or the DNA construct comprising said nucleotide sequence.50. The cell of claim 49 , wherein the cell is a bacterial cell claim 49 , archaeal cell claim 49 , fungal cell claim 49 , animal cell or plant cell.51. A method of driving a reaction which produces PPi towards the production of PPi claim 44 , the method comprising: adding the HPPA of to the reaction mixture of the reaction which produces PPi.52. The method of claim 51 , wherein the reaction is an enzymatic reaction.53. The method of claim 52 , wherein the enzymatic reaction is performed under high temperature.54. The method of claim 52 , wherein the enzymatic reaction is performed ...

Electrochemical measurement method, electrochemical measurement device and transducer

An electrochemical measurement method for electrochemically measuring a chemical substance generated or consumed in a biological sample in a solution is provided which includes performing measurement by placing the biological sample at a distance away from an electrode surface in the direction perpendicular to the electrode surface. The distance is determined in advance on the basis of simulation in which a current flowing through a working electrode.

MASS SPECTROMETRY METHOD FOR MEASURING THIAMINE IN BODY FLUID

Provided are methods for determining the amount of total thiamine in a body fluid sample using liquid chromatography and mass spectrometry. Total thiamine is converted to free thiamine by treatment with an acid phosphatase prior to thiamine separation and quantification. 116.-. (canceled)17. A method for determining the amount of thiamine in a sample , comprising:(i) purifying thiamine from the sample by high performance liquid chromatography (HPLC);(iii) ionizing the purified thiamine by electrospray ionization (ESI);(iv) determining the amount of thiamine by mass spectrometry, wherein the amount of total thiamine in the sample is determined.18. The method of claim 17 , the method further comprising removing soluble protein from a plasma or serum sample;19. The method of claim 18 , wherein soluble protein is removed by treating said sample with an acid.20. The method of claim 17 , the method further comprising incubating the sample with an acid phosphatase claim 17 , for not longer than about 2 hours to convert phosphorylated thiamine to thiamine;21. The method of claim 20 , wherein the incubation is carried out at a pH of about 4.6±0.1.22. The method of claim 20 , wherein the incubation is carried out at a temperature of about 40° C.23. The method of claim 20 , wherein the incubation is carried out between about 1 and about 2 hours.24. The method of claim 17 , wherein the liquid chromatography comprises high performance liquid chromatography (HPLC).25. The method of claim 17 , wherein the ionizing comprises ionizing a parent ion having a mass/charge ratio of 265.00±1.0.26. The method of claim 25 , wherein the parent ion is fragmented into one or more daughter ions and wherein the amount of one or more of said daughter ions is determined.27. The method of claim 26 , wherein the one or more daughter ions comprises an ion having a mass/charge ratio of 144.00±1.0 or 121.94±1.0.28. The method of claim 26 , wherein the one or more daughter ions comprise an ion having a ...

METHOD FOR ANALYZING DNA METHYLATION USING NEXT GENERATION SEQUENCER AND METHOD FOR CONCENTRATING SPECIFIC DNA FRAGMENTS

This invention provides a technology of DNA methylation analysis including: 1. A method for determining a methylation state in a DNA to be analyzed , with said method comprising of:(1) digesting the DNA to be analyzed with a restriction enzyme whose recognition site includes methylated cytosine or cytosine to be methylated and is affected by methylation;(2) ligating a mixture of DNA fragments obtained in step (1) with a ligase;(3) determining a nucleotide sequence of each DNA construct included in a mixture of DNA constructs obtained in step (2); and(4) comparing a nucleotide sequence of each recognition site of the restriction enzyme and its flanking nucleotide sequences included in each nucleotide sequence information obtained in step (3) with a known genomic sequence, and determining whether each recognition site is a recognition site that has not been digested with the restriction enzyme, or a recognition site that has been digested with the restriction enzyme and regenerated by the ligation with the ligase, to thereby determine the methylation state at each recognition site.2. The method according to claim 1 , wherein the mixture of DNA fragments obtained in step (1) is ligated with the ligase in the presence of an adaptor capable of being ligated to its both ends claim 1 , in step (2).3. The method according to claim 1 , wherein a desired DNA fragment group is fractionated from the mixture of DNA fragments obtained in step (1) before the ligation with the ligase claim 1 , in step (2).4. The method according to claim 1 , wherein a DNA amplification is carried out using a DNA polymerase with strand displacement activity after the ligation with the ligase claim 1 , in step (2).5. The method according to claim 1 , wherein a nucleotide sequence between adjacent recognition sites of the restriction enzyme is mapped to a known genomic sequence claim 1 , and a sequence outside at least one of the adjacent recognition sites is compared with the mapped reference ...

Protecting groups for biological labeling

Alpha-haloketones are useful alkylating agents for coupling to sulfhydryl-containing biomolecules. However, they react spontaneously with water, alkali and organic bases and therefore cannot be stored for extended periods of time in aqueous solutions, particularly in the presence of proteins at physiological pH. The present invention provides novel solutions to these problems, however, as novel compounds and compositions comprising protected haloketones are disclosed herein. Methods of preparing and using protected haloketones which are useful in a variety of applications—e.g., in assays and conjugation reactions—are also disclosed herein.

Combination for target marker detection

본 발명은 표적 마커를 간편하고 고감도로 검출하기 위한 조합물에 관한 것이다. 보다 상세하게는, 본 발명은 표적 마커와 특이적으로 결합 가능한 표적 마커 결합 분자와 결합하여 생물학적 시료 중에서 표적 마커를 검출하기 위한 조합물로서 (a) 상기 표적 마커 결합 분자와 직접 또는 간접적으로 특이적으로 결합 가능한 제1 결합 분자와 표지 물질을 포함하는 제1 결합제; (b) 상기 제1 결합제와 특이적으로 결합 가능한 링커 분자; 및 (c) 상기 링커 분자와 특이적으로 결합할 수있는 제2 결합제이며, 제2 결합 분자와 표지 물질을 포함하는 제2 결합제를 적어도 포함하는 조합물에 관한 것이다. The present invention relates to a combination for simple and highly sensitive detection of a target marker. More specifically, the present invention is a combination for detecting a target marker in a biological sample by binding with a target marker-binding molecule capable of specifically binding to a target marker. (a) a first binding agent comprising a first binding molecule and a labeling substance capable of specifically binding directly or indirectly to the target marker binding molecule; (b) a linker molecule capable of specifically binding to the first binding agent; And (c) a second binding agent capable of specifically binding to the linker molecule, and relates to a combination comprising at least a second binding agent including a second binding molecule and a labeling substance.

Patent RU2016145600A3

РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2016 145 600 A (51) МПК G01N 33/536 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ЗАЯВКА НА ИЗОБРЕТЕНИЕ (21)(22) Заявка: 2016145600, 23.04.2015 (71) Заявитель(и): НИТИРЕЙ БАЙОСАЙЕНСИЗ ИНК. (JP) Приоритет(ы): (30) Конвенционный приоритет: 23.04.2014 JP 2014-089675 (85) Дата начала рассмотрения заявки PCT на национальной фазе: 23.11.2016 R U (43) Дата публикации заявки: 25.05.2018 Бюл. № 15 (72) Автор(ы): ОБАЯСИ Хирокадзу (JP), ФУДЗИТА Каеко (JP) (86) Заявка PCT: (87) Публикация заявки PCT: WO 2015/163424 (29.10.2015) R U (54) КОМБИНИРОВАННЫЙ ПРОДУКТ ДЛЯ ДЕТЕКЦИИ МАРКЕРА-МИШЕНИ (57) Формула изобретения 1. Комбинированный продукт для детекции маркера-мишени в биологическом образце в комбинации с молекулой, связывающейся с маркером-мишенью, способной специфически связываться с маркером-мишенью, содержащий, по меньшей мере: (a) первое связывающее средство, содержащее первую связывающую молекулу, способную прямо или косвенно специфически связываться с молекулой, связывающейся с маркером-мишенью, и вещество мечения; (b) линкерную молекулу, способную специфически связываться с первым связывающим средством; и (c) второе связывающее средство, способное специфически связываться с линкерной молекулой и содержащее вторую связывающую молекулу и вещество мечения. 2. Комбинированный продукт по п. 1, где линкерная молекула способна специфически связываться с веществом мечения. 3. Комбинированный продукт по п. 1 или 2, где линкерная молекула представляет собой антитело или его антигенсвязывающий фрагмент. 4. Комбинированный продукт по любому из пп. 1-3, где вещество мечения представляет собой по меньшей мере одно выбранное из хемилюминесцентной метки, металлической частицы, флуоресцентной метки, ферментной метки, коферментной метки, меченного антитела, красителя, биолюминесцентной метки, гаптена и полимерной частицы. Стр.: 1 A 2 0 1 6 1 4 5 6 0 0 A Адрес для переписки: 129090, Москва, ул. Б. Спасская, 25, стр. 3, ООО " ...

Комбинированный продукт для детекции маркера-мишени

РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2016 145 600 A (51) МПК G01N 33/536 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ЗАЯВКА НА ИЗОБРЕТЕНИЕ (21)(22) Заявка: 2016145600, 23.04.2015 (71) Заявитель(и): НИТИРЕЙ БАЙОСАЙЕНСИЗ ИНК. (JP) Приоритет(ы): (30) Конвенционный приоритет: 23.04.2014 JP 2014-089675 (85) Дата начала рассмотрения заявки PCT на национальной фазе: 23.11.2016 R U (43) Дата публикации заявки: 25.05.2018 Бюл. № 15 (72) Автор(ы): ОБАЯСИ Хирокадзу (JP), ФУДЗИТА Каеко (JP) (86) Заявка PCT: (87) Публикация заявки PCT: WO 2015/163424 (29.10.2015) R U (54) КОМБИНИРОВАННЫЙ ПРОДУКТ ДЛЯ ДЕТЕКЦИИ МАРКЕРА-МИШЕНИ (57) Формула изобретения 1. Комбинированный продукт для детекции маркера-мишени в биологическом образце в комбинации с молекулой, связывающейся с маркером-мишенью, способной специфически связываться с маркером-мишенью, содержащий, по меньшей мере: (a) первое связывающее средство, содержащее первую связывающую молекулу, способную прямо или косвенно специфически связываться с молекулой, связывающейся с маркером-мишенью, и вещество мечения; (b) линкерную молекулу, способную специфически связываться с первым связывающим средством; и (c) второе связывающее средство, способное специфически связываться с линкерной молекулой и содержащее вторую связывающую молекулу и вещество мечения. 2. Комбинированный продукт по п. 1, где линкерная молекула способна специфически связываться с веществом мечения. 3. Комбинированный продукт по п. 1 или 2, где линкерная молекула представляет собой антитело или его антигенсвязывающий фрагмент. 4. Комбинированный продукт по любому из пп. 1-3, где вещество мечения представляет собой по меньшей мере одно выбранное из хемилюминесцентной метки, металлической частицы, флуоресцентной метки, ферментной метки, коферментной метки, меченного антитела, красителя, биолюминесцентной метки, гаптена и полимерной частицы. Стр.: 1 A 2 0 1 6 1 4 5 6 0 0 A Адрес для переписки: 129090, Москва, ул. Б. Спасская, 25, стр. 3, ООО " ...

혈액 중 간 효소 수준을 측정하기 위한 시스템 및 방법

혈액 샘플 중 하나 이상의 간 효소의 농도를 측정하기 위한 일회용 검사 카트리지는 적어도 하나의 챔버가 혈액 샘플 중 간 효소 ALT, AST, ALP, 또는 GGT 중 하나에 의해 촉매화될 때 반응하여 반응 용액을 형성하는 반응물 혼합물을 함유하는 복수의 챔버를 갖는 카트리지 바디, 카트리지 바디에 연결된 탈착식 카트리지 커버, 및 카트리지 바디에 연결된 검사 스트립 모듈을 포함한다. 검사 스트립 모듈은 반응 용액의 일부를 수용하고 반응 용액에서 반응 생성물인 적어도 하나의 분석물을 전기화학적으로 측정하도록 구성된 적어도 하나의 분석물 검사 스트립을 포함한다.

Method for determining a ratio of RNA type in a sample

원형 RNA를 이용하여 시료 중 RNA로부터 DNA를 생성하는 방법을 제공한다.

Methods, products, and kits for identifying an analyte in a sample

Methods, products, and kits for identifying an analyte in a sample are provided. One embodiment of a method for identifying an analyte in a sample includes combining the sample with a first reactant capable of specifically coupling to the analyte. The first reactant is coupled to beads. The method also includes combining additional reactant with the beads. The additional reactant is capable of specifically coupling to the analyte or a second reactant coupled to an analyte. An enzyme is attached to the additional reactant. In addition, the method includes combining a substrate with the beads. The substrate is capable of specifically interacting with the enzyme to form a modified substrate. If the substrate interacts with the enzyme attached to the beads via the additional reactant, the solubility of the substrate changes causing the modified substrate to bind to a surface of the beads and/or the reactants bound to the beads. The method further includes identifying the analyte in the sample by detecting the modified substrate bound to the surface of the beads and/or the reactants bound to the beads.

Cap-pap test

The CAP-PAP Test is a double-staining, single-slide microscopic method. An in vitro diagnostic medical device for manual and automatic staining and interpreting of the Pap smear for cervical cancer screening, cervical dysplasia and for follow-up therapy can be developed using this double-staining, single-slide microscopic method. Abnormal cervical cells are labeled with an intracellular acid phosphatase derived pigment (azo-dye) to improve visibility of abnormal cervical cells on conventionally stained Pap smears. The enzyme marker improves human perception and/or sensitivity of automatic instruments when distinguishing cell a abnormality and interpretation of Pap smears. Increased accuracy of CAP-PAP-vs-Pap test is expected to reduce false negative readings of the conventional Pap test. A rapid manual version of the test that is low cost, does not require additional personnel training and is instantly applicable in all cytopathology laboratories is provided. The invention further provides a diagnostic kit, an automatic stainer and an automatic evaluation device for performing the double-staining, single-slide microscopic method.

Method of substances and their compositions detection with anticancer activity

FIELD: medicine. SUBSTANCE: invention relates to development of drug preparations for oncological diseases treatment. Disclosed is method for detection of substances and their compounds with anticancer activity, based on reporter protein production increasing coded by recombinant replicative-defective adenovirus, in response to substances or their compositions exposure on molecular targets from protein kinase mTOR, topoisomerases I and II, histone deacetylase and unknown targets, inhibited by LY294002 and LY303511 compositions. EFFECT: invention enables efficient detection of substances having anticancer activity, and compositions thereof. 10 cl, 4 dwg, 4 ex РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (51) МПК G01N 33/52 (13) 2 602 452 C2 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ (21)(22) Заявка: ИЗОБРЕТЕНИЯ К ПАТЕНТУ 2014119354/15, 14.05.2014 (24) Дата начала отсчета срока действия патента: 14.05.2014 (43) Дата публикации заявки: 20.11.2015 Бюл. № 32 (73) Патентообладатель(и): Федеральное государственное бюджетное учреждение науки Институт биологии гена Российской академии наук (RU) R U Приоритет(ы): (22) Дата подачи заявки: 14.05.2014 (72) Автор(ы): Шепелев Михаил Валентинович (RU), Коробко Елена Владимировна (RU), Коробко Игорь Викторович (RU) (45) Опубликовано: 20.11.2016 Бюл. № 32 C 2 2 6 0 2 4 5 2 R U (54) СПОСОБ ВЫЯВЛЕНИЯ ВЕЩЕСТВ И ИХ КОМПОЗИЦИЙ С ПРОТИВООПУХОЛЕВОЙ АКТИВНОСТЬЮ (57) Реферат: Изобретение относится к области разработки веществ или их композиций на молекулярные лекарственных препаратов для лечения мишени из числа протеинкиназы mTOR, онкологических заболеваний. Предложен способ топоизомераз I и II, гистон деацетилаз и выявления веществ и их композиций с неизвестных мишеней, ингибируемых противоопухолевой активностью, основанный соединениями LY294002 и LY303511. Изобретение на увеличении продукции репортерного белка, обеспечивает эффективное выявление веществ, кодируемого рекомбинантным репликативнообладающих ...

Combination for target marker detection

FIELD: measurement; testing.SUBSTANCE: group of inventions relates to detection of target marker in a biological sample. Kit for detecting a target marker in a biological sample in combination with a molecule, binding to the target marker, capable of specific binding to the target marker includes: first binding agent comprising a first binding molecule capable of directly or indirectly specifically binding to the molecule binding to the target marker and the labelling substance; linker molecule capable of specific binding to the first binding agent; and a second binding agent comprising a second binding molecule capable of specific binding to said linker molecule and a target substance. Wherein the labelling substance in the first binding agent and the labelling substance in the second binding agent may be the same or different; linker molecule is capable of specifically binding to the labelling substance in the first binding agent and the labelling substance in the second binding agent; first binding agent is a structure in which the first binding molecule and the labelling substance are connected to each other directly or indirectly by means of a carrier, which is a natural or synthetic polymer; second binding agent is a structure in which the second binding molecule and the labelling substance are connected to each other directly or indirectly by means of a carrier which is a natural or synthetic polymer. Method for detecting a target marker in a biological sample and the use of a kit for detecting a target marker in a biological sample are also disclosed.EFFECT: group of inventions provides high detection sensitivity of the target marker.11 cl, 11 dwg, 7 tbl РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 678 108 C2 (51) МПК G01N 33/536 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (52) СПК G01N 33/536 (2018.08) (21)(22) Заявка: 2016145600, 23.04.2015 (24) Дата начала отсчета срока действия патента: (73) ...

Measuring method of electrochemical biosensor

본 발명은 전기화학신호 측정원리로 혈액, 배뇨, 타액 등 체액에 포함된 바이오마커를 검출하는 전기화학식 바이오 센서의 측정방법에 관한 것으로, 더욱 상세하게는 전기화학식 바이오 센서를 이용한 체외 진단기기에서 시간전류법 또는 시간전하법으로 측정함에 있어서 측정의 정확성 및 재현성을 개선할 수 있는 전기화학식 바이오 센서의 측정방법에 관한 것이다. 본 발명에 따르면, 바이오마커를 측정하기 위한 전기화학식 바이오 센서를 시간전류법 또는 시간전하법으로 측정함에 있어서, 전극을 측정용액에 담그고 일정시간 전극에 전압을 걸어주지 않은 후 전하값을 측정하거나 일정시간 전압을 걸어준 후 전류값을 측정함으로써 측정의 정확성 및 재현성을 개선할 수 있는 우수한 효과가 있다. The present invention relates to a measuring method of an electrochemical biosensor that detects biomarkers contained in body fluids such as blood, urination, saliva, etc. using the electrochemical signal measurement principle. It relates to a measurement method of an electrochemical biosensor capable of improving the accuracy and reproducibility of measurement in measurement by an amperometric method or a time charge method. According to the present invention, in measuring an electrochemical biosensor for measuring a biomarker by the time amperometric method or the time charge method, the electrode is immersed in a measurement solution and a voltage is not applied to the electrode for a certain period of time, and then the charge value is measured or There is an excellent effect of improving the accuracy and reproducibility of measurement by measuring the current value after applying a time voltage.

Method of enhancing the efficacy of anti-hepatocyte growth factor receptor breast cancer therapy by administering an inhibitor of menaINV

Methods and compositions are provided for diagnosing or inhibiting invasion or metastasis of a cancer in a subject based on Mena INV .

Kinase and phosphatase assays

Compositions, methods, and kits for detecting and monitoring kinase, phosphatase and protein post-translational modification activity are described. The compositions typically include a peptide, a detectable moiety, and a protease cleavage site. Modification of a peptide by a kinase, phosphatase or other protein post-translational modification alters the proteolytic sensitivity of the peptide, resulting in a change of a detectable property of the composition. Panel assays for determining substrates or modulators of kinase, phosphatase or other protein post-translational modification activity are also described.

Method for improving alkaline phosphatase assay

The present invention relates to a method for improving an ALP assay. More specifically, the present invention relates to a method that can be used for an ALP assay immediately after ATDC5 cell freezing and thawing regardless of the storage period, without requiring a long incubation period, by specifying the freezing conditions of ATDC5 cells to provide. Accordingly, it is possible by the present invention to more rapidly and easily measure osteoblast differentiation activity of BMP protein by performing ALP assay.

Reagents for the detection of protein phosphorylation in carcinoma signaling pathways

The invention discloses nearly 474 novel phosphorylation sites identified in signal transduction proteins and pathways underlying human carcinoma, and provides phosphorylation-site specific antibodies and heavy-isotope labeled peptides (AQUA peptides) for the selective detection and quantification of these phosphorylated sites/proteins, as well as methods of using the reagents for such purpose. Among the phosphorylation sites identified are sites occurring in the following protein types: Kinase, Adaptor/Scaffold proteins, Phosphatase, G protein Regulator/Guanine Nucleotide Exchange Factors/GTPase Activating Proteins, Cytoskeleton Proteins, DNA Binding Proteins, Phospholipase, Receptor Proteins, Enzymes, DNA Repair/Replication Proteins, Adhesion Proteins, and Proteases , as well as other protein types.

Bimolecular optical probes

Compositions, methods, and kits for detecting and monitoring post-translational modification activities, including kinase or phosphatase activities, are described. The compositions typically include a peptide, a first detectable moiety, a first binding member, and a protease cleavage site. Modification of a composition by a post-translational modification enzyme, such as a kinase or phosphatase, alters the proteolytic sensitivity of the peptide, resulting in a change of a detectable property of the composition when it is associated noncovalently with a probe composition that includes a second binding member and a second detectable moiety. Panel assays for determining substrates or modulators of enzymatic activities are also described.

Assay for determining inhibitors of ATPase

This invention provides materials and methods for identifying inhibitors of a Hepatitis C Virus NS3 protein ATPase. Methods for making and purifying such an ATPase are also provided by this invention.

Sample preparation method for DNA methylation analysis

Sampling devices and methods of use

Combination product for testing goal marker

本发明涉及一种用于简易且高灵敏度地检测目的标志物的组合产品。更详细而言，本发明涉及一种用于与目的标志物结合分子并用而检测生物学试样中的目的标志物的组合产品，上述目的标志物结合分子能够与生物学试样中的目的标志物特异性结合，该组合产品至少包含：(a)第一结合剂，其包含能够与上述目的标志物结合分子直接或间接地特异性结合的第一结合分子、和标记物质；(b)能够与上述第一结合剂特异性结合的连接分子；以及(c)第二结合剂，其为能够与上述连接分子特异性结合的第二结合剂，包含第二结合分子和标记物质。

Molecular interactions in cells

The invention provides reagents and methods for inhibiting or enhancing interactions between proteins in cells, particularly interactions between a PDZ protein and a PL protein. Reagents and methods that are provided are useful for treatment of a variety of diseases and conditions in a variety of cell types.

Room-temperature phosphorescence detection method of alkaline phosphatase and application

一种碱性磷酸酶的室温磷光检测方法及应用，属于碱性磷酸酶的检测技术领域，可解决现有碱性磷酸酶的检测过程复杂，成本高以及干扰大的问题。室温下，本发明利用制备的β‑环糊精修饰的Mn:ZnS室温磷光量子点为磷光探针，焦磷酸根为分子识别单元，通过检测加入碱性磷酸酶后体系的室温磷光变化实现对焦磷酸酶的分析检测。该磷光检测体系对焦磷酸酶的响应范围为0.22‑10.4 U/L，检出限为0.045 U/L。可用于血清中碱性磷酸酶的检测，且在测定时无需复杂的样品预处理过程。室温磷光的产生也不需要加入除氧剂和诱导剂，并能够避免实际样品本底荧光和散射光的干扰。

Simple and rapid acid phosphatase activity determination method

本发明公开了一种简单快捷的酸性磷酸酶活性测定方法，具体步骤如下：步骤一，取得一条链的3`端磷酸化的双链DNA；步骤二，以该双链DNA为底物在酸性磷酸酶存在下在反应体系中反应，将反应获得的具有3`端去磷酸化的双链DNA的反应物在足量的核酸外切酶存在下反应；步骤三，反应完成后，测定反应体系中单链DNA和双链DNA的相对量，并由该检测结果推导出酸性磷酸酶的活性程度。本发明公布了一种简单快捷的酸性磷酸酶活性测定方法，该方法比现有技术操作步骤更简单、方便和快捷，可实现高通量和自动化的活性测定，并且灵敏度高，可检测到15mU的酸性磷酸酶，使用效果好。

Electrochemical measurement method, electrochemical measurement apparatus and transducer

【課題】溶液中の生体サンプルで生成または消費される化学物質を電気化学的に測定する電気化学測定において、測定感度の向上を図る方法を提供する。【解決手段】化学物質と電子の授受を行なって酸化還元反応をさせる作用極の電極面の寸法と、生体サンプルの寸法とを与件に含み、生体サンプルにおいて化学物質が生成または消費される化学反応の速度と、溶液中の化学物質の濃度分布の変化と、電極面における化学物質との電子の授受の速度とに基いて作用極に流れる電流を計算するシミュレーションを、生体サンプルと電極面との電極面に対する垂直方向距離を変数として行って、電流が最大値となる点を含み電流が最大値の９０％以上の大きさとなる垂直方向距離の範囲を求め、溶液中において生体サンプルを、電極面から電極面に対する垂直方向に前記範囲に属する距離だけ離間させて測定を実行する。【選択図】なし A method for improving measurement sensitivity in an electrochemical measurement in which a chemical substance generated or consumed in a biological sample in a solution is electrochemically measured. A chemistry in which a chemical substance is generated or consumed in a biological sample, including a dimension of an electrode surface of a working electrode that exchanges electrons with the chemical substance to cause an oxidation-reduction reaction and a dimension of the biological sample. A simulation that calculates the current that flows to the working electrode based on the reaction rate, the change in concentration distribution of the chemical substance in the solution, and the rate of transfer of electrons with the chemical substance on the electrode surface, The vertical distance to the electrode surface is determined as a variable, and the range of the vertical distance including the point where the current is the maximum value and the current is 90% or more of the maximum value is obtained. Measurement is performed by separating the surface from the surface by a distance belonging to the range in a direction perpendicular to the electrode surface. [Selection figure] None

Monolithic multilayered analytical element for measuring activity of alkaline phosphatase

(57)【要約】本公報は電子出願前の出願データであるた め要約のデータは記録されません。

Methods for measuring enzyme activity useful in determining cell viability in non-purified samples

METHOD FOR IDENTIFICATION OF SUBSTANCES AND THEIR COMPOSITIONS WITH ANTITUMOR ACTIVITY (OPTIONS)

1. Способ выявления веществ и их композиций с противоопухолевой активностью, предусматривающий следующие стадии:а) рассев клеток;б) заражение клеток рекомбинантным репликативно-дефектным аденовирусом, кодирующим репортерный белок;в) инкубацию зараженных клеток для наработки начального уровня репортерного белка;г) добавление к клеткам испытуемого вещества или их композицией (опыт) или растворителя испытуемого вещества или их композиции (контроль);д) инкубацию опытных и контрольных клеток;е) определение уровня продукции репортерного белка в опытных и контрольных клетках;ж) определение количества опытных и контрольных клеток;з) нормирование уровней репортерного белка в опыте и контроле на соответствующее количество клеток;е) сравнение уровней репортерного белка в опыте и контроле, причем увеличение уровня репортерного белка в опыте по сравнению с контролем означает, что испытуемое вещество или их композиция оказала воздействие на одну или несколько мишеней в клетках из числа протеинкиназы mTOR, топоизомераз I и II, гистон деацетилаз и неизвестных мишеней, в частности, ингибируемых соединениями LY294002 и LY303511, и обладает потенциальной противоопухолевой активностью.2. Способ по п. 1, в котором клетки являются опухолевыми клетками человека или других млекопитающих.3. Способ по п. 1, в котором клетки рассеиваются и анализ проводится в многолуночном, предпочтительно в 96-луночном, планшете.4. Способ по п. 1, в котором в качестве репортерного белка, кодируемого рекомбинантным репликативно-дефектным аденовирусом, используется зеленый флуоресцентный белок EGFP.5. Способ по п. 1, в котором уровень экспрессии репортерного гена � РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (51) МПК G01N 33/52 (13) 2014 119 354 A (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ЗАЯВКА НА ИЗОБРЕТЕНИЕ (21)(22) Заявка: 2014119354/15, 14.05.2014 (71) Заявитель(и): Федеральное государственное учреждение науки Институт биологии гена Российской академии наук (RU) Приоритет(ы): (22) Дата ...

Adipocyte differentiation detecting marker and a method for regulating differentiation of mesenchymal stem cells using same

본 발명은 PTPRM (protein tyrosin phosphatase receptor type M), PTPRF (protein tyrosin phosphatase receptor type F), PTPRQ (protein tyrosin phosphatase receptor type Q), SSH2 (slingshot homolog 2) 및 MTMR11 (myotubularin related protein 11)로 이루어진 군으로부터 선택되는 단백질 타이로신 포스파타제(PTP)의 발현수준을 측정하는 제제를 포함하는, 인간 중간엽줄기세포로부터 지방세포로의 분화 여부를 탐지하기 위한 마커 검출용 조성물, 상기 조성물을 포함하는 키트, 상기 마커 단백질을 이용하여 지방세포로의 분화를 탐지하는 방법, 지방세포로의 분화 조절 후보 화합물을 스크리닝하는 방법, 및 지방세포로의 분화를 조절하는 방법에 관한 것이다. The present invention relates to a group consisting of protein tyrosin phosphatase receptor type M (PTPRM), protein tyrosin phosphatase receptor type F (PTPRF), protein tyrosin phosphatase receptor type Q (PTPRQ), slingshot homolog 2 (SSH2) and myotubularin related protein 11 (MTMR11). Marker detection composition for detecting the differentiation of human mesenchymal stem cells into adipocytes, comprising a preparation for measuring the expression level of protein tyrosine phosphatase (PTP) selected from, a kit comprising the composition, the marker protein The present invention relates to a method for detecting differentiation into adipocytes, a method for screening differentiation candidate compounds into adipocytes, and a method for controlling differentiation into adipocytes. 중간엽줄기세포, 탈인산화효소, 리버스 트랜스크립타제 피씨알 Mesenchymal stem cells, dephosphatase, reverse transcriptase PCR