Determinationofthetiter
Todeterminethetiter,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainsthumanJMJD2c(Cat.#25298).Thewellswerecoatedwiththepeptideusedforimmunisationoftherabbit.Byplottingtheabsorbanceagainsttheantibodydilution(Figure1),thetiteroftheantibodywasestimatedtobe1:2,000.
WesternblotanalysisusingtheantibodydirectedagainstJMJD2c
NuclearextractsofHeLacells(40µg)wereanalyzedbyWesternblotusingtheantibodyagainstJMJD2c(Cat.#25298)diluted1:1,000inTBS-Tweencontaining5%skimmedmilk.Thepositionoftheproteinofinterest(expectedsize:120kDa)isindicatedontheright;theMarker(inkDa)isshownontheleft.Thesmallerfragmentofapproximately92kDamayrepresentasplicingvariant.
Storeat–80°Cforupto2years.Centrifugeafterfirstthawtomaximizeproductrecovery.Aliquottoavoidrepeatedfreeze/thawcycles.Aliquotsmaybestoredat–20°Cforatleastonemonth.
WB(1:1000)