Background 
5-hydroxymethylcytosine(5-hmC),asasixthDNAbasewithfunctionsintranscriptionregulation,hasbeendetectedtobeabundantinhumanandmousebrainandembryonicstem(ES)cells.Inmammals,itcanbegeneratedbyoxidationof5-methylcytosine(5-mC),areactionmediatedbytheten-eleventranslocation(TET)familyof5-mChydroxylases.5-hmCwasdemonstratedtoplayanimportantanddifferentrolethan5-mCintheregulationofDNAmethylation,chromatinremodeling,andgeneexpression.

Concentration 
1mg/ml

Description 
Mousemonoclonalantibodyto5-Hydroxymethylcytosine(5-hmC),cloneHMC/4D9

Specificity 
Modifiedbase5-hydroxymethylcytosine(5-hmC),abroadrangeofspecies

Isotype 
IgG1

Formulation 
10mMphosphatebuffer,150mMNaCl,pH7.4 

Storage  
4°C.Forlong-termstorage,aliquotandstoreat-20°C.Avoidrepeatedfreezingandthawing.Multiplefreeze/thawcyclesmayresultindecreasedperformance.Stablefor6monthsfromthedateofshipment.

Purity 
ProteinApurified

HandlingRecommendations 
Formaximumrecoveryoftheproducts,centrifugethevialpriortoopeningthecap

AlternativeNames 
5-hydroxymethylcytidine,anti-5-hydroxymethylcytidine,anti-5-hydroxymethylcytosine,anti-5hmC,anti-5-hmC,anti-5-hmeC,anti-5hmeC,5hmC,5hmeC,5-hmeC 

Application 
Immunofluorescence:1:200–1:500
Immunohistochemistry:1:200–1:500
ELISA:1:1000–1:2000
DotBlot: 1:2000
hMeDIP:0.5-1µg/10⁶Cells

Fig.1.DotblotanalysisofA-1018withDNAoligoscontainingcytosine(C),5-methylcytosine(5mC),or5-hydroxymethylcytosine(5hmC).10ngofC,5mC,and5hmCDNAoligoseachwerespottedonamembrane.Themembranewasincubatedwith0.5μg/mlofEpigentek"s5hmCCloneHMC/4D9antibody(1:2000dilution),thenwithperoxidaseconjugatedgoatanti-mouseIgG(dilution1:3000).Specificsignalwasonlyobservedwith5-hmCDNAoligos.


Fig.2.DemonstrationofhighsensitivityandspecificityofEpigentek"s5-hmCCloneHMC/4D9antibodyagainst5-hmCDNA.SyntheticDNAoligoscontaining5-hmC,5-mCoronlycytosine,respectivelywereaddedintotheassaywellsatdifferentconcentrationsandthenspecificsignalswasonlydetectedwith5-hmCDNA.

Fig.3. SelectiveenrichmentofhydroxymethyatedDNA.50pgofunmethylated,methylated,andhydroxymethylatedDNAoligoswereeachspikedintofragmentedhumangenomicDNA(500ng).hMeDIPwasprocessedwithEpigentek"s5-hmCCloneHMC/4D9antibody.ElutedDNAwasanalyzedbyrealtimePCRwiththeprimerstodetectthepresenceofspikedDNAoligos.Fold-enrichmentrepresentstheamountofrecoveredDNAoligosandwascalculatedbasedontheCts.