ApplicationNotes | Westernblot:1-10μg/mL
CellpelletsareresUSPendedin100μLlysisbuffer(25mMTris,pH7.6,150mMNaCl,1μg/mLaprotonin,10μg/mLleupeptin,1mMphenylmethylsulfonylfluoride,1mMsodiumorthovanadate,1mMEDTA,and1%TritonX-100).Sampleswereincubatedfor1hourat4°Candwerepelletedathighspeedfor10minutesat4°C,and100μgproteinpersamplewassubjectedto10%sodiumdodecylsulfate-polyacrylamidegelelectrophoresis.
ELISA:1:100,000primaryEIAusingimmunogenonly(50-100ngSERTpeptide(CatalogNumberAG297)/well).Nottestedonpurifiedproteinatthestateddilutions.
Optimalworkingdilutionsmustbedeterminedbyenduser. |