Applications
ApplicationNotes
ValidatedApplications:
DB:0.5-1µg/mldilution
Immunogen
This5-Carboxylcytosineantibodywasraisedagainst5-carboxylcytidineconjugatedtoKLHandrecognizes5-carboxylcytosine.
Buffer
PurifiedIgGinPBSwith30%glyceroland0.035%sodiumazide.Sodiumazideishighlytoxic.
References
Dotblotof5-CarboxylcytosinepAb.
Dotblotanalysiswasusedtoconfirmthespecificityof5-Carboxylcytosineantibodyfor5-Carboxylcytosine(5-carboxylcytidine,5-caC).Single-strandedDNAoligonucleotides(amountofoligoinnanogramslistedontheleftsideoftheblot)correspondingtotheimmunogenandrelatedsequenceswerespottedontonitrocelluloseandprobedwiththeantibodyat0.5µg/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