ChIPresultsobtainedwiththeantibodydirectedagainstH2Apan
ChIPassayswereperformedusinghumanHeLacells,theantibodyagainstH2Apan(Cat.#25226)andoptimizedPCRprimersetsforqPCR.ChIPwasperformedonshearedchromatinfrom10,000cellsusingtheSX-8GIP-Starautomatedsystem.Atitrationoftheantibodyconsistingof1,2,5,and10µgperChIPexperimentwasanalyzed.IgG(5µg/IP)wasusedasnegativeIPcontrol.QPCRwasperformedwithprimersfortheGAPDHpromoterandfortheinactiveMYODgene.Figure1showstherecovery,expressedasa%ofinput(therelativeamountofimmunoprecipitatedDNAcomparedtoinputDNAafterqPCRanalysis).

Determinationofthetiter
Todeterminethetiteroftheantibody,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainstH2Apan(Cat.#25226),crudeserumandflow-throughinantigencoatedwells.Byplottingtheabsorbanceagainsttheantibodydilution(Figure2),thetiteroftheantibodywasestimatedtobe1:1,800.

CrossreactivitytestusingtheantibodydirectedagainstH2Apan
ADotBlotanalysiswasperformedtotestthecrossreactivityoftheantibodyagainstH2Apan(Cat.#25226)withthepeptideusedforimmunizationoftherabbitandotherpeptidescontainingunmodifiedsequencesofdifferenthistones.Onehundredto0.2pmoloftherespectivepeptideswerespottedonamembrane.Theantibodywasusedatadilutionof1:20,000.Figure3showsahighspecificityoftheantibodyfortheH2Apeptide.

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