首页 > Active Motif > Active Motif（艾跃）/5-Hydroxymethylcytosine (5-hmC) antibody (pAb)/39770/10µl

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Active Motif（艾跃）/5-Hydroxymethylcytosine (5-hmC) antibody (pAb)/39770/10µl

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39770

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ApplicationsApplicationNotesValidatedApplications:MeDIP:0.1-0.5µlperIPDB:1:10,000dilutionPublishedApplications:MeDIPMeDIP-SeqICC/IFIHC(FFPE)FCDBSeereferencesformoreinformation.Individualoptimizationmayberequired.ImmunogenThis5-Hydroxymethylcytosineantibodywasraisedagainst5-hydroxymethylcytidineconjugatedtoKLHandrecognizes5-hydroxymethylcytosine.BufferRabbitserumcontaining30%glyceroland0.035%sodiumazide.Sodiumazideishighlytoxic.References5-Hydroxymethylcytosine(5-hmC,5-hydroxymethylcytidine)antibodytestedbyMethylDNAimmunoprecipitation.DNA(25pg)derivedfromthepromoteroftheAPCgenewasspikedinto500ngofhumangenomicDNAandsubjectedtotheMeDIPprocedureusing1μlof5-Hydroxymethylcytidineantibody(5hmC,maroonbars)or1μlofcontrolrabbitIgG(IgG,bluebars).RealtimequantitativePCRwasperformedontheimmunoprecipitatedDNAandresultsplottedas%ofinputDNA.ThespikedAPCDNAcontainedeithernomethylation(DNA),5-methylcytosinemethylation(5-mC)or5-hydroxymethylcytosinemethylation(5-hmC).5-Hydroxymethylcytosine(5-hmC,5-hydroxymethylcytidine)antibodytestedbydotblotanalysis.DNAsamples(10ngor50ngasindicated)werespottedontopositivelychargednylonmembraneandblottedwith5-Hydroxymethylcytidineantibodyatadilutionof1:10,000.TopPanel:DoublestrandedDNA.BottomPanel:SinglestrandedDNA.Lane1:UnmethylatedDNA.Lane2:DNAcontaining5-methylcytosine.Lane3:DNAcontaining5-hydroxymethylcytosine.5-Hydroxymethylcytosine(5-hmC,5-hydroxymethylcytidine)antibodytestedbydotblotanalysis.DNAsampleswerespottedontopositivelychargednylonmembraneandblottedwithantibodiesasindicated.PanelA:5-Hydroxymethylcytidineantibodyrecognizing5-hydroxymethylcytosine(1:10,000dilution).PanelB:5-Methylcytidineantibody(1:1,000dilution).Lane1:DNAderivedfrommouseembryonicstemcells(150ng).Lane2:DNAderivedfrommousespleen(600ng).Lane3:27baseoligonucleotidecontaining5-hydroxymethylcytosine(1.2ng).Lane4:33baseoligonucleotidecontaining5-methylcytosine(2000ng).BackgroundActiveMotifofferstwopolyclonalantibodiesthatrecognize5-hydroxymethylcytosine,awholeserumversion(39769)andapurifiedIgGversion(39791).BotharevalidatedforuseinmethylDNAimmunoprecipitation(MeDIP).ForcustomerswhomustquantitatetheamountofIgGintheMeDIPreaction,thepurifiedIgGversion(39791)isrecommended.Thewholeserumversion(39769)isveryhightitre,soshouldbeusedcarefully(0.1-0.5µlperIP)topreventnon-specificbackground.Thewholeserumversion(39769)hasbeenusedsuccessfullyinimmunofluorescence(IF,Itoetal,2010);thepurifiedIgGversion(39791)islikelytoworkinIFaswell.DNAmethylationisanepigeneticeventinwhichDNAmethyltransferases(DNMTs)catalyzethereactionofamethylgrouptothefifthcarbonofcytosineinaCpGdinucleotide.Thismodificationhelpstocontrolgeneexpressionandisalsoinvolvedingenomicimprinting,whileaberrantDNAmethylationisoftenassociatedwithdisease.5-methylcytosineisamodifiedbasethatisfoundintheDNAofplantsandvertebrates.AsecondtypeofDNAmethylationexists,5-hydroxymethylcytosine(5-hydroxymethylcytosine,5-hmC).Thisresultsfromtheenzymaticconversionof5-methylcytosineinto5-hydroxymethylcytosinebytheTETfamilyofcytosineoxygenases.Thisantibodywasdevelopedspecificallytodistinguish5-hydroxymethylcytosinefrom5-methylcytosineasconventionalmethods(enrichmentbyantibodyormethylDNAbindingprotein,enzymaticdigestionandbisulfitesequencing)cannotdoso.ItispossIBLethat5-hydroxymethylcytosine(5-hmC)representsapathwaytodemethylateDNA,as5-hydroxymethylcytosineisrepairedasmismatchedDNAandreplacedwithunmethylatedcytosine.CompatibleSecondaryAntibodiesActiveMotifhasdevelopedavarietyofhigh-qualitysecondaryantibodiesformostapplications,withanti-rabbitandanti-mouseconjugatestoabroadlineofhigh-qualityfluorescentmolecules,aswellashorserADIshperoxidase(HRP).VisitourSecondaryAntibodyConjugatespagetofindtheperfectsecondariesforimmunofluorescenceexperimentswith5-Hydroxymethylcytosine(5-hmC)antibody(pAb).StorageAntibodiesinsolutioncanbestoredat-20°Cfor2years.Avoidrepeatedfreeze/thawcyclesandkeeponicewhennotinstorage.GuaranteeThisproductisguaranteedfor6monthsfromdateofreceipt.Thisproductisforresearchuseonlyandisnotforuseindiagnosticprocedures.ApplicationKeyChIP=ChromatinImmunoprecipitation;DB=DotBlot;ELISA=Enzyme-linkedImmunosorbentAssay;EMSA=ElectrophoreticMobilityShiftAssayFC=FlowCytometry;ICC=Immunocytochemistry;IF=Immunofluorescence;IHC=Immunohistochemistry;IP=Immunoprecipitation;MeDIP=Methyl-DNAImmunoprecipitation;TR-FRET=Time-ResolvedFluorescenceResonanceEnergyTransfer;WB=WesternBlotting

Active Motif 公司是一家致力于研发和生产核细胞分子生物学和表观遗传学研究用试剂的专业公司，也是世界上最大的转录因子试剂生产商和 ChIP-seq 外包服务供应商。Active Motif 的产品涉及：表观遗传学相关抗体、Chip 级别抗体、重组识别子和催化酶、预甲基化修饰的重组组蛋白、多聚核小体和生物素标记核小体；以及各类染色质分析、转录因子表达调控分析和 DNA 甲基化分析的试剂盒等。不仅如此，Active Motif 还为研究者提供专业的表观遗传学服务，特别是 Chip-seq 的外包服务。公司总部位于美国加州，目前在欧洲，日本，中国设有分公司。产品销售范围覆盖全球 30 多个国家和地区。