Rabbit anti bax and anti caspase 3 antibodies

We analyzed HEI-OC1 cell extracts by 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis. We electro-transferred the separated proteins to nitrocellulose membranes, blocked the membranes with a solution of 20 mM Tris-HCl (pH 7.6), 137 mM NaCl, and 0.01% Tween-20 (TBS-T) containing 5% skim milk for 1 h, and then probed the membranes with primary antibodies (1:100–1:2000 dilution) at RT. Next, we washed the membranes three times for 5 min each with TBS-T and incubated them with secondary antibody (1:2000 dilution). After a series of washes, we developed the membranes using an enhanced chemiluminescent detection system. The primary antibodies were rabbit anti-Bax and anti-caspase-3 antibodies from Cell Signaling (Danvers, MA, USA) and rabbit anti-Bcl-2 from Abcam (Cambridge, MA, UK). The secondary antibody was goat anti-rabbit-IgG-HRP (Cell Signaling).

HEI-OC1 cell extracts were analyzed by 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis. The separated proteins were electrotransferred to a nitrocellulose membrane. Membranes were blocked with a solution of 20 mM Tris-HCl (pH 7.6), 137 mM NaCl, and 0.01% Tween-20 (TBS-T) containing 5% skim milk for 1 h, and then probed with primary antibodies (1:100–1:2000 dilution) at room temperature. Next, the membranes were washed three times for 5 min each with TBS-T and incubated with secondary antibody (1:2000 dilution). After a series of washes, the membranes were developed using an enhanced chemiluminescent detection system. Rabbit anti-Bax and anti–caspase-3 antibodies were from Cell Signaling (Cell Signaling, Danvers, MA, USA), and rabbit anti-Bcl-2, anti-GR, and anti-GPx antibodies were from Abcam (Abcam, Cambridge, MA, UK). Goat anti-rabbit-lgG-HRP was used as the secondary antibody (Cell Signaling, Danvers, MA, USA).