Antibodies against akt and p akt

GSRd was obtained from Tai-He Biopharmaceutical (Guangzhou, China). MPP⁺ iodile, MPTP, α-tocopherol and LY294002 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Hoechst 33342, rhodamine 123 (Rh123), 2,7-dichlorofluorescein diacetate (DCFH-DA) were purchased from Beyotime Institute of Biotechnology (Shanghai, China). The commercial kits for detection of lipid peroxidation MDA, SOD, GPX, and ATP bioluminescence were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The commercial kit for mitochondrial respiration complex I activity detection was purchased from Genmed Scientifics Corp (Shanghai, China). Antibodies against TH, Bax and Bcl-2 were purchased from Abcam (Cambridge, UK), and antibodies against Akt and P- Akt were purchased from Cell Signaling (Carlsbad, CA, USA). Cy2 conjugated anti-rabbit IgG were purchased from Vector laboratory (Burlingame, CA, USA).

Equal amounts of cellular proteins were resolved by electrophoresis in 12% (SDS-PAGE) and transferred to a nitrocellulose membrane for immunoblotting using specific antibodies. Antibodies against HER2 and PARP were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against AKT and P-AKT were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA); antibodies against CDK4, CDK6, Caspase 3 and LAMP-1 were purchased from Genetex (Taiwan); antibodies against PSMD3 were purchased from Bethyl Laboratory (Montgomery, AL, USA). The β-actin was used as an internal control and purchased from Sigma (Montgomery, AL, USA). Primary antibodies were incubated at 4 °C overnight, followed by incubation with secondary antibodies (goat anti-mouse or goat anti-rabbit) for an hour at room temperature. The antigen-antibody complexes were detected using the Western Blotting Luminol Reagent Kit (Santa Cruz, CA, USA).