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Caspase 3 Antibody (35-1600Z) in WB
Western blot analysis of Caspase-3 was performed by loading 20 ug of U-87 MG (lane 1), Hep G2 (lane 2), Ntera-2 (lane 3), Jurkat (lane 4) and HEK-293 (lane 5) (Fig. A) and Jurkat (lane 1), Jurkat treated for O/N with 3 uM of Staurosporine (lane 2), HeLa (lane 3), HeLa treated for O/N with 1 uM of Etoposide (lane 4) and HeLa treated for O/N with 3 uM of Staurosporine in (lane 5) (Fig. B) cell lysates using Novex® NuPAGE® 4-12 % Bis-Tris gel (NP0322BOX), XCell SureLock? Electrophoresis System (EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. Caspase-3 was detected at ~32 kDa using Caspase-3 Mouse Monoclonal Antibody (351600Z) at 1-2 µg/mL in 5 % skim milk at 4°C overnight on a rocking platform. Goat Anti-Mouse - HRP Secondary Antibody (626520) at 1:4000 dilution was used and chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (WP20005). (Fig. B) Upon treatment with Staurosporine/Etoposide there was a reduction in total Caspase-3.
