Anti bcl xl ab32370

Tissues were homogenized and sonicated in radio-immunoprecip itation assay lysis buffer (Millipore, Billerica, USA), supplemented with protease and phosphatase inhibitors (Roche Diagnostics, Basel, Switzerland). Proteins (80 μg) were mixed with loading buffer and denatured by boiling for 5 min. The proteins were separated by 4%-13% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then transferred to a nitrocellulose membrane (Millipore, Billerica, USA). After being blocked with 5% non-fat milk, membranes were then incubated with primary antibodies overnight at 4°C, followed by incubation with horseradish peroxidase-conjugated secondary antibodies at room temperature for 1 h. Finally, protein bands were visualized using and enhanced chemiluminescence detection kit (Millipore). The primary antibodies used were: anti-GAPDH (ab8245), anti-beta actin (ab8226), anti-HGF (ab83760), anti-cleaved caspase 3 (ab2302), and caspase 9 (ab2324), antiapoptosis signal-regulating kinase (anti-ASK)-1 (ab131506), antiphospho-JNK (ab47337), anti-Bax (ab32503), anti-Bcl-2 (ab59348), and anti-Bcl-xl (ab32370) antibodies, which were all from Abcam, and anti-cleaved caspase 8 (#9496) antibody which was from Cell Signaling Technology (Danvers, USA).