ChIPresultsobtainedwiththeantibodydirectedagainstH2Bpan
ChIPassayswereperformedusinghumanHeLacells,theantibodyagainstH2Bpan(Cat.#25231)andoptimizedPCRprimersetsforqPCR.ChIPwasperformedonshearedchromatinfrom10,000cells.Atitrationoftheantibodyconsistingof1,2,5,and10µgperChIPexperimentwasanalyzed.IgG(5µg/IP)wasusedasnegativeIPcontrol.QPCRwasperformedwithprimersfortheGAPDHpromoterandfortheinactiveMYODgene.Figure1showstherecovery,expressedasa%ofinput(therelativeamountofimmunoprecipitatedDNAcomparedtoinputDNAafterqPCRanalysis).

Determinationofthetiter
Todeterminethetiteroftheantibody,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainstH2Bpan(Cat.#25231),crudeserumandflow-throughinantigencoatedwells.Byplottingtheabsorbanceagainsttheantibodydilution(Figure2),thetiteroftheantibodywasestimatedtobe1:14,950.

CrossreactivitytestusingtheantibodydirectedagainstH2Bpan
ADotBlotanalysiswasperformedtotestthecrossreactivityoftheantibodyagainstH2Bpan(Cat.#25231)withthepeptideusedforimmunizationoftherabbitandotherpeptidescontainingunmodifiedsequencesofdifferenthistones.Onehundredto0.2pmoloftherespectivepeptideswerespottedonamembrane.Theantibodywasusedatadilutionof1:20,000.Figure3showsahighspecificityoftheantibodyfortheH2Bpeptide. 

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