A - Western blot analysis of Caspase 3 expression in rat liver (1), rat thymus (2) and SMCC whole cell lysate. Caspase-3 is detected at the expected molecular weight of 31 kDa. Samples were loaded at 50 µg/lane, and SDS-PAGE performed under reducing conditions. Transfer: nitrocellulose membrane. Blocking: 5% skim milk, 1.5 h at RT. Primary antibody: 0.5 µg/mL overnight at 2-8°C. Secondary antibody: Goat anti-rabbit IgG-HRP (1:10,000), 1.5 h at RT. Detection: ECL.
B - Analysis of Caspase 3 expression in paraffin-embedded section of human lung cancer tissue by Immunohistochemistry. Heat-mediated antigen retrieval was performed in citrate buffer (pH6) for 20 minutes. The tissue section was blocked with 10% goat serum and then incubated with 1 µg/mL rabbit anti-caspase 3 antibody overnight at 2-8°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using DAB as the chromogen.