Determinationofthetiter
Todeterminethetiter,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainstmouseSetd1b(Cat.No.25314).Theplateswerecoatedwithamixofthepeptidesusedforimmunizationoftherabbit.Byplottingtheabsorbanceagainsttheantibodydilution(Figure1),thetiteroftheantibodywasestimatedtobe1:8,400.
WesternblotanalysisusingtheantibodydirectedagainstmouseSetd1b
Westernblotwasperformedonwholecelllysatesfrommousefibroblasts(NIH3T3)andembryonicstemcells(E14Tg2a)withtheantibodyagainstmouseSetd1b(Cat.No.25314),diluted1:500inBSA/PBS-Tween.ThemolecularweightMarker(inkDa)isshownontheleft;thelocationoftheproteinofinterestisindicatedontheright.
Storeat–80°Cforupto2years.Centrifugeafterfirstthawtomaximizeproductrecovery.Aliquottoavoidrepeatedfreeze/thawcycles.Aliquotsmaybestoredat–20°Cforatleastonemonth.
WB(1:500)