Determinationofthetiter
Todeterminethetiter,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainstmouseG9a(Cat.No.25224).Thewellswerecoatedwiththepeptidesusedforimmunisationoftherabbit.Byplottingtheabsorbanceagainsttheantibodydilution(Figure1),thetiteroftheantibodywasestimatedtobe1:30,000.
WesternblotanalysisusingtheantibodydirectedagainstG9a
Westernblotwasperformedonwholecelllysatesfrommousefibroblasts(NIH3T3)andembryonicstemcells(E14Tg2a)cellswiththeantibodyagainstmouseG9a(Cat.No.25224),diluted1:1,000inBSA/PBS-Tween.ThemolecularweightMarker(M,inkDa)isshownontheleft;thelocationoftheproteinofinterest(138kDa)isindicatedontheright.
Storeat–80°Cforupto2years.Centrifugeafterfirstthawtomaximizeproductrecovery.Aliquottoavoidrepeatedfreeze/thawcycles.Aliquotsmaybestoredat–20°Cforatleastonemonth.
WB(1:1000)
