首页 > Active Motif > Active Motif（艾跃）/5-Hydroxymethylcytosine (5-hmC) antibody (rAb)/91309/50µg

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Active Motif（艾跃）/5-Hydroxymethylcytosine (5-hmC) antibody (rAb)/91309/50µg

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ApplicationsApplicationNotesValidatedApplications:DB:0.2-1µg/mlIHC:0.1-1µg/mlICC:0.5-2µg/mlMeDIP:0.2-2µg/mlImmunogenBSA-conjugated5-hydroxymethylcytosine.BufferPBSwith50%glycerol,1%BSA,and0.09%sodiumazide.ReferencesJ.Reichmannetal."Dualspindleformationinzygoteskeepsparentalgenomesapartinearlymammalianembryos."bioRxiv(2017)10.1101/198275.Siref,A.B.etal."Diagnosticutilityofdual5-hmC/Melan-Aimmunohistochemistryindifferentiatingnodalnevusfrommetastaticmelanoma:Aneffectivefirst-linetestforthework-upofsentinellymphnodespecimen."J.CutanPathol.(2019)10.1111/cup.13412.Dotblotof5-Hydroxymethylcytosine(5-hmC)antibody(rAb).Membranewaspre-spottedwith50,5,and0.5ng/dotofdoublestranded5-Hydroxymethylcytosine(5-hmC)DNA,5-Methylcytosine (5-mC)DNA,andunmethylatedDNA.Thepre-spottedmembranewasthenblottedwithantibodyat0.2µg/ml.MeDIPof5-Hydroxymethylcytosine(5-hmC)antibody(rAb).MeDIPwasperformedusing5-Hydroxymethylcytosineantibody(CloneRM236)ata10:1DNA:Abratio.1ngofunmethylated,5-Methylcytosine(5-mC)or5-Hydroxymethylcytosine(5-hmC)DNAstandard(897bp)wasspikedin1µgof genomicDNAisolatedfromHeLacellsasthecontrol.RealtimePCRwasthenperformedtodeterminethecaptureofDNAstandardasin%ofinput. ImmunofluoresencestainingofHeLacellsusing5-Hydroxymethylcytosineantibody(CloneRM236)at0.5µg/mL(red).Actinfilamentsarelabeledwithfluoresceinphalloidin(green).HeLacellswerefixedwith4%paraformaldehydeandpermeABIlizedwithmethanol(−20°C)beforetreatmentwith2NHClfor30minat37°CtodenatureDNA.Immunohistochemicalstainingofformalinfixed,paraffinembeddedhumanbraintissuesections,using5-Hydroxymethylcytosineantibody(CloneRM236)at1µg/ml.BackgroundDNAmethylationisanepigeneticeventinwhichDNAmethyltransferases(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(rAb).StorageSomeproductsmaybeshippedatroomtemperature.Thiswillnotaffecttheirstabilityorperformance.Uponreceipt,unconjugatedantibodiesmaybestoredat-20°Cforupto2years.Fluorophore-&enzyme-conjugatedantibodiesshouldbestoredat4°C.Fluorophore-conjugatedantibodiesshouldbeprotectedfromlight.Keepreagentsonicewhennotinstorage;toavoidrepeatedfreeze/thawcycles,werecommendaliquotingitemsthatwillbestoredfrozenintosingle-usefractionspriortofreezing.GuaranteeThisproductisforresearchuseonlyandisnotforuseindiagnosticprocedures.Thisproductisguaranteedfor6monthsfromdateofarrival.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。