Applications

Western Blot ValidatedDot Blot Validated

ApplicationNotes


ValidatedApplications:
DB:0.5-2µg/mldilution

PublishedApplications:
WB
Seereferencesformoreinformation.Individualoptimizationmayberequired.

Immunogen

This5-Formylcytosineantibodywasraisedagainst5-formylcytidineconjugatedtoKLHandrecognizes5-formylcytosine.

Buffer

PurifiedIgGinPBSwith30%glyceroland0.035%sodiumazide.Sodiumazideishighlytoxic.

References

5-Formylcytosine (5-fC) antibody (pAb) tested by dot blot analysis.

Dotblotof5-FormylcytosinepAb.
Dotblotanalysiswasusedtoconfirmthespecificityof5-Formylcytosineantibodyfor5-Formylcytosine(5-formylcytidine,5-fC).Single-stranded38ntDNAoligonucleotides(amountofoligoinnanogramslistedontheleftsideoftheblot)correspondingtotheimmunogenandrelatedsequenceswerespottedontonitrocelluloseandprobedwiththeantibodyat0.2µg/ml.Lane1:oligocontaining5-carboxylcytidine.Lane2:oligocontaining5-formylcytidine.Lane3:oligocontaining5-hydroxymethylcytidine.Lane4:oligocontaining5-methylcytidine.Lane5:oligocontainingunmodifiedcytidine.

Background

DNAmethylationisanepigeneticeventinwhichDNAmethyltransferases(DNMTs)catalyzethereactionofamethylgrouptothefifthcarbonofcytosineinaCpGdinucleotide.Thismodificationhelpstocontrolgeneexpressionandisalsoinvolvedingenomicimprinting,whileaberrantDNAmethylationisoftenassociatedwithdisease.5-methylcytosineisamodifiedbasethatisfoundintheDNAofplantsandvertebrates.

AsecondtypeofDNAmethylationexists,5-hydroxymethylcytosine(5-hydroxymethylcytosine,5-hmC).Thisresultsfromtheenzymaticconversionof5-methylcytosineinto5-hydroxymethylcytosinebytheTETfamilyofcytosineoxygenases.IterativeactivityofTETon5-hydroxymethylcytosineresultsintheconversionof5-hmCinto5-formylcytosineand5-carboxylcytosine.Anincreaseinlevelsof5-formylcytosineand5-carboxylcytosinearedetectedinthemousemalepronucleusfollowingfertilization,whichisgraduallydilutedbyDNAreplication.

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