Determinationofthetiter
Todeterminethetiter,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainstH3K18ac(Cat.No.25236).Theantigenusedwasapeptidecontainingthehistonemodificationofinterest.Byplottingtheabsorbanceagainsttheantibodydilution(Figure1),thetiteroftheantibodywasestimatedtobe1:8,800.

CrossreactivitytestusingtheantibodydirectedagainstH3K18ac
ADotBlotanalysiswasperformedtotestthecrossreactivityoftheantibodyagainstH3K18ac(Cat.No.25236)withwithpeptidescontainingothermodificationsandunmodifiedsequencesofhistoneH3.Onehundredto0.2pmolofthepeptidecontainingtherespectivehistonemodificationwerespottedonamembrane.Theantibodywasusedatadilutionof1:1,000.Figure2showsahighspecificityoftheantibodyforthemodificationofinterest.

WesternblotanalysisusingtheantibodydirectedagainstH3K18ac
HeLacellsweretreatedwithbutyrateandhistoneextracts(15μg)wereanalysedbyWesternblotusingtheantibodyagainstH3K18ac(Cat.No.25236)diluted1:250inTBS-Tweencontaining5%skimmedmilk(lane1).Lane2showsacoomassiebluestainingofthegel.Thepositionoftheproteinofinterestisindicatedontheleft;theMarker(inkDa)isshownontheright.

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