Determinationoftheantibodytiter
Todeterminethetiteroftheantibody,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainsthumanDNMT3A(Cat.#25218),crudeserumandflow-throughinantigencoatedwells.Byplottingtheabsorbanceagainsttheantibodydilution(Figure1),thetiteroftheantibodywasestimatedtobe1:2,000.

WesternblotandImmunoprecipitationusingtheantibodydirectedagainstDNMT3A
Humanembryonickidneycells(HEK293T)weretransientlytransfectedwithanexpressionvectorforGAL4-taggedDNMT3A.Wholecellextractswereimmunoprecipitatedwith2µgofantibodyagainstDNMT3A(Cat.#25218).ThepresenceofGAL4-DNMT3Ainthenon-treatedcellextractsandintheimmunoprecipitateswasdemonstratedbyWesternblotanalysiswithanti-GAL4antibody(lane1and2respectively).Alternatively,GAL4-DNMT3AwasimmunoprecipitatedandWesternblotanalysiswasperformedwiththeDNMT3Aantibody(diluted1:2,500)(lane3).

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