InputType: | NuclearExtracts,PurifiedEnzyme |
ResearchArea: | DNAMethylation |
TargetApplication: | ActivityMeasurement |
VesselFormat: | 96-WellPlate |
100%Guarantee: | 6months |
TheEpigenase™ 5mC-HydroxylaseTETActivity/InhibitionAssayKit(Colorimetric)isacompletesetofoptimizedbuffersandreagentsformeasuringactivity/inhibitionoftotalcytosineoxygenaseTETenzymesinnuclearextractsorpurifiedTETisoforms(TETs1-3)fromabroadrangeofspeciessuchasmammals,plants,fungi,andbacteria,andinavarietyofformsincluding,butnotlimitedtoculturedcellsandfreshandfrozentissues. Thekithasthefollowingadvantagesandfeatures:
BackgroundInformation
5-hydroxymethylcytosine,asasixthDNAbasewithfunctionsintranscriptionregulation,hasbeendetectedtobeabundantinthehumanandmousebrainandembryonicstem(ES)cells.Inmammals,itcanbegeneratedbyoxidationof5-methylcytosine,areactionmediatedbytheten-eleventranslocation(TET)familyofcytosineoxygenases.
TheTETfamilyofcytosineoxygenasesincludesTET1,TET2,andTET3.TheseTETproteinsmaypromoteDNAdemethylationbybindingtoCpG-richregionstopreventunwanted DNAmethyltransferase activity,andbyconverting5mCto5hmCandfurtherto5caC(5-carboxylcytosine)throughhydroxylaseactivity.Itwasshownthatgenomic5hmClevelscorrelatewithTEThydroxylaseactivity.Inaddition, TET1wasshowntohavedualfunctionsintranscriptionactivationandrepressionbybindingdifferenttargetgenesinEScells.TET1isalsoafusionpartneroftheMLLgeneinacutemyeloidleukemiaandisconsideredanoncoprotein.TET2isfoundtobefrequentlymutatedinleukemiaandconsideredtoactastumorsuppressor.TET3wasdemonstratedtoplayauniqueroleinDNAmethylationreprogrammingprocessesinthemammalianzygote.Thus,activatingtumorsuppressorTETenzymessuchasTET2orinhibitingoncoproteinTETenzymessuchasTET1wouldbeimportantinbenefitingcancerdiagnositcsanddevelopingnewtarget-basedcancertherapeutics.
Principle&Procedure
Inthisassay,amethylatedsubstrateisstablycoatedontothemicroplatewells.ActiveTETs(notincluded)bindtothesubstrateandconvertthemethylatedsubstratetohydroxymethylatedproducts.TheTET-convertedhydroxymethylatedproductscanberecognizedwithaspecificantibody.Theratiooramountofhydroxymethylatedproducts,whichisproportionaltoenzymeactivity,canthenbecolorimetricallymeasuredbyreADIngtheabsorbanceinamicroplatespectrophotometeratawavelengthof450nm.TheactivityoftheTETenzymeisinturnproportionaltotheopticaldensityintensitymeasured.
WB(10XWashBuffer)
TAB(TETAssayBuffer)
TS(10XTETSubstrate)*
BS(BindingSolution)
HC5(TETAssayStandard,20µg/ml)*
HC6(CaptureAntibody,1000µg/ml*
HC7(DetectionAntibody,400µg/ml)*
HC8(EnhancerSolution)*
DS(DeveloperSolution)
SS(StopSolution)
Co-factor1*
Co-factor2*
Co-factor3*
8-WellAssayStrips(WithFrame)
*Formaximumrecoveryoftheproducts,centrifugetheoriginalvialpriortoopeningthecap.
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作为表观遗传学公司,Epigentek Group Inc.是表观遗传相关研究的创新技术和产品的领先开发商和提供商。自2005年成立以来,该公司已经开发了一系列专有产品,可为表观遗传学相关的研究和药物发现提供完整,系统的解决方案。
Epigentek产品在全球享有盛誉的学术界,领先的制药公司和尖端生物技术公司广泛使用。这些独特的产品经过专门设计,可以使检测比当前使用的方法更简单,更快,更方便且更高效。Epigentek的产品通过遍布36个国家和地区的专业销售团队和全球分销商网络进行销售。确保我们技术的创新将使客户完全满意并在您的实验中获得最大的成功是我们的第一要务。
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