ChIPresultsobtainedwiththemonoclonalantibodydirectedagainstH3K9me3
ChIPassayswereperformedonhumanHeLacellsusingthemonclonalantibodyagainstH3K9me3(cat.No.25271).ChIPwasperformedusingshearedchromatinfrom1,000,000cells.Atitrationconsistingof1,2,5and10μgofantibodyperChIPexperimentwasanalyzed.IgG(2μg/IP)wasusedasanegativeIPcontrol.QPCRwasperformedwithprimersforthepromotersoftheactivec-fosandGAPDHgenes,usedasnegativecontrols,andfortheZNF510geneandtheSat2satelliterepeatregion,usedaspositivecontrols.Figure1showstherecovery,expressedasa%ofinput(therelativeamountofimmunoprecipitatedDNAcomparedtoinputDNAafterqPCRanalysis).

CrossreactivitytestusingthemonoclonalantibodydirectedagainstH3K9me3
ADotBlotanalysiswasperformedtotestthecrossreactivityofthemonoclonalantibodyagainstH3K9me3(cat.No.25271)withpeptidescontainingdifferentmodificationsofhistoneH3orH4andtheunmodifiedH3K9sequence.Onehundredto0.2pmolofpeptidecontainingtherespectivehistonemodificationwerespottedonamembrane.Theantibodywasusedatadilutionof1:100,000.Figure2showsahighspecificityoftheantibodyforthemodificationofinterest.

WesternblotanalysisusingthemonoclonalantibodydirectedagainstH3K9me3
Westernblotwasperformedonhistoneextracts(15μg,lane1)fromHeLacells,andon1μgofrecombinanthistoneH3(lane2)usingthemonoclonalantibodyagainstH3K9me3(cat.No.25271).Theantibodywasdiluted1:1,000inTBS-Tweencontaining5%skimmedmilk.TheMarker(inkDa)isshownontheleft,thepositionoftheproteinisindicatedontheright.

Storeat–80°Cforupto2years.Centrifugeafterfirstthawtomaximizeproductrecovery.Aliquottoavoidrepeatedfreeze/thawcycles.Aliquotsmaybestoredat–20°Cforatleastonemonth.