ChIPresultsobtainedwiththeantibodydirectedagainstH3K4ac
ChIPassayswereperformedusingWTandH3K4RmutantS.pombecells,theantibodyagainstH3K4ac(Cat.#25251)andoptimizedprimerpairsforqPCR.Shearedchromatincorrespondingto10µgofDNAand0.5µgofantibodywereusedperChIPexperiment.QPCRwasperformedusingprimersspecificfortwodifferentpericentricrepeatregionsandfortheeuchromaticadh1gene.Figure1showstherecovery,expressedasa%ofinput(therelativeamountofimmunoprecipitatedDNAcomparedtoinputDNA).

Determinationoftheantibodytiter
Todeterminethetiteroftheantibody,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainstH3K4ac(Cat.#25251)inantigencoatedwells.Theantigenusedwasapeptidecontainingthehistonemodificationofinterest.Byplottingtheabsorbanceagainsttheantibodydilution(Figure2),thetiterofthepurifiedantibodywasestimatedtobe1:27,800.

CrossreactivitytestusingtheantibodydirectedagainstH3K4ac
ADotBlotanalysiswasperformedtotestthecrossreactivityoftheantibodyagainstH3K4ac(Cat.#25251)withpeptidescontainingotherhistoneH3modificationsandtheunmodifiedH3K4sequence.Onehundredto0.2pmoloftherespectivepeptideswerespottedonamembrane.Theantibodywasusedatadilutionof1:10,000.Figure3showsahighspecificityoftheantibodyforthemodificationofinterest.

WesternblotanalysisusingtheantibodydirectedagainstH3K4ac
Histoneextracts(15µg)fromHeLacellswereanalyzedbyWesternblotusingtheantibodydirectedagainstH3K4ac(Cat.#25251),diluted1:500inTBS-Tweencontaining5%BSA.TheMarker(inkDa)isshownontheleft,thepositionoftheproteinofinterestisindicatedontheright.

ImmunofluorescencewiththeantibodydirectedagainstH3K4ac
WildtypeandH3K4RmutantS.pombecellswerestainedwithboththeantibodyagainstH3K4ac(Cat.#25251)(inred)andbyHoechststaining(inblue,left),orwiththeH3K4acantibodyalone(right).Theantibodywasusedatadilutionof1:300. 

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