Applications

Immunofluorescence ValidatedDot Blot ValidatedImmunocytochemistry Validated

ApplicationNotes


ValidatedApplications:
DB:0.1-1µg/mldilution

PublishedApplications:
ICC/IF
DotBlot
Pleaseseereferencesformoreinformation.Individualoptimizationmayberequired.

Immunogen

This3-methylcytosineantibodywasraisedagainst3-methylcytidineconjugatedtoKLHandrecognizes3-methylcytosine.

Buffer

PurifiedrabbitIgGinPBSwith30%glyceroland0.035%sodiumazide.Sodiumazideishighlytoxic.

References

3-Methylcytosine (3-mC) antibody (pAb) tested by dot blot analysis.

3-Methylcytosine(3-mC,3-methylcytidine)antibodytestedbydotblotanalysis.
BSAconjugatednucleosides(startingat10ngasindicated)werespottedontoPVDFmembraneandblottedwith3-methylcytidineantibodyatadilutionof1:10,000.
Lane1:3-methylcytidine.
Lane2:Cytidine.
Lane3:5-methylcytidine.
Lane4:5-hydroxymethylcytidine.

Background

MethylationofDNAcanoccurnon-enzymaticallyatthenitrogen-threeofthecytosinebasethroughspontaneousexposuretoendogenousS-adenosylmethionine(SAM).Theresulting3-methylcytosine(3-mC)ismutagenicandmustberepaired,whichoccursinhumansthroughthebaseexcisionrepair(BER)ordealkylationviahumanhomologuesoftheE.coliAlkBprotein.3-methylcytosineispresentinhumancelllinesandincreasedlevelsof3-mCimpairproliferation.

CompatIBLeSecondaryAntibodies

ActiveMotifhasdevelopedavarietyofhigh-qualitysecondaryantibodiesformostapplications,withanti-rabbitandanti-mouseconjugatestoabroadlineofhigh-qualityfluorescentmolecules,aswellashorserADIshperoxidase(HRP).VisitourSecondaryAntibodyConjugatespagetofindtheperfectsecondariesforimmunofluorescenceexperimentswith3-Methylcytosine(3-mC)antibody(pAb).

Storage

Someproductsmaybeshippedatroomtemperature.ThiswillnotaffecttheirstABIlityorperformance.Uponreceipt,unconjugatedantibodiesmaybestoredat-20°Cforupto2years.Fluorophore-&enzyme-conjugatedantibodiesshouldbestoredat4°C.Fluorophore-conjugatedantibodiesshouldbeprotectedfromlight.Keepreagentsonicewhennotinstorage;toavoidrepeatedfreeze/thawcycles,werecommendaliquotingitemsthatwillbestoredfrozenintosingle-usefractionspriortofreezing.

Guarantee

Thisproductisguaranteedfor6monthsfromdateofreceipt.

Thisproductisforresearchuseonlyandisnotforuseindiagnosticprocedures.

ApplicationKey

  • ChIP=ChromatinImmunoprecipitation;
  • DB=DotBlot;
  • ELISA=Enzyme-linkedImmunosorbentAssay;
  • EMSA=ElectrophoreticMobilityShiftAssay
  • FC=FlowCytometry;
  • ICC=Immunocytochemistry;
  • IF=Immunofluorescence;
  • IHC=Immunohistochemistry;
  • IP=Immunoprecipitation;
  • MeDIP=Methyl-DNAImmunoprecipitation;
  • TR-FRET=Time-ResolvedFluorescenceResonanceEnergyTransfer;
  • WB=WesternBlotting