Western Blot Analysis of Bcl-2, Bax, Cytochrome C, Caspase-9, Caspase-3 Expression

Cells were treated with baicalein at a concentration of 50 and 100 µM of baicalein for 24 h. Equal protein concentrations of lysate were separated by 12% SDS-PAGE and then transferred into PVDF membranes. The membrane was pre-incubated with TBS buffer containing 5% BSA and 0.05% Tween 20 for 2 h at room temperature and then blotted with a primary monoclonal antibody (Bcl-2, Bax, Caspase-9, Caspase-3, and β-actin) overnight at 4°C, followed by membrane were incubated with appropriate horseradish peroxidase-conjugated secondary antibodies for 1 h. Protein bands were detected by an enhanced chemiluminescence detection kit.