Lane 1 = Bax protein (Tagged) (ab85157) – 10 ng. Lane 2 = Extract of HeLa cells – 40 ug. Lane 3 = Extract of HepG2 cells – 40 ug. Lane 4 = Bax protein (Tagged) (ab85157) – 10 ng. Lane 5 = Extract of HeLa cells – 40 ug. Lane 6 = Extract of HepG2 cells – 40 ug. SDS PAGE performed under reducing conditions (100mM DTT – Sample heated at 50ºC). Primary : Lanes 1-3: Anti Bax antibody ( ab77566 ) at 1 ug/mL. Lanes 4-6: Anti Bax antibody ( ab32503 ) at 1:2000 dilution. Secondary : Lanes 1-3: Goat anti mouse IgG(H&L)-HRP at 1:10000. Lanes 4-6: Goat anti rabbit IgG(H&L)-HRP at 1:10000. Development: ECL with 2 min exposure. Blocking: in 5% Milk + PBS for 3 hours at RT. Primary antibody: in 5% Milk + PBS overnight at 4 C. Secondary antibody: in 5% Milk + PBS for 2 hour at RT. Predicted band size : Bax 21kDa and Bax (Tagged) 49 kDa. Observed band size : Bax 21kDa and Bax (Tagged) 49 kDa
SDS-PAGE showing ab85158 at approximately 49kDa.
Standard curve for Bax (Analyte: ab85157 ); dilution range 1pg/ml to 1µg/ml using Capture Antibody ab77566 at 0.2µg/ml and Detector Antibody ab32503 at 0.5µg/ml. Concentration of ab32503 may vary from lot to lot; please use this curve as guideline.