Determinationofthetiter
Todeterminethetiter,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainstmouseSetd1a(Cat.#25313).Theplateswerecoatedwithamixofthepeptidesusedforimmunizationoftherabbit.Byplottingtheabsorbanceagainsttheantibodydilution(Figure1),thetiteroftheantibodywasestimatedtobe1:2,500.
WesternblotanalysisusingtheantibodydirectedagainstmouseSetd1a
Westernblotwasperformedonwholecelllysatesfrommousefibroblasts(NIH3T3)andembryonicstemcells(E14Tg2a)withtheantibodyagainstmouseSetd1a(Cat.#25313),diluted1:500inBSA/PBS-Tween.ThemolecularweightMarker(inkDa)isshownontheleft;thelocationoftheproteinofinterest(expectedsize:186kDa)isindicatedontheright.
Storeat–80°Cforupto2years.Centrifugeafterfirstthawtomaximizeproductrecovery.Aliquottoavoidrepeatedfreeze/thawcycles.Aliquotsmaybestoredat–20°Cforatleastonemonth.
WB(1:500)