Bcl 2 rabbit antibody

To investigate the presence of apoptotic signals, tissue of the right temporal lobe of sampled mice brains was homogenized in RIPA lysis buffer (Beyotime) and Pierce Protease and Phosphatase Inhibitor Mini Tablets (Thermo Scientific), and centrifuged (13,000 g for 15 minutes at 4°C). Protein concentrations were determined using BCA Protein Assay Kits (Thermo Fisher Scientific). An equal amount of protein samples (40 μg/lane) were loaded onto a 12% SDS-PAGE and transferred onto PVDF membranes (Merck Millipore). The membranes were blocked with 5% milk powder and incubated overnight at 4°C with the primary antibodies: Bcl-2 Rabbit Antibody (1:1000, Cell Signaling Technology #3498) and Bax Rabbit Antibody (1:1000, Cell Signaling Technology #2772) and β-actin Rabbit Antibody (1:10000, Cell Signaling Technology #4970). The PVDF membranes were then incubated with the HRP-linked secondary antibody (1:5000, Santa Cruz Biotechnology) at RT for 1 h. Protein bands were visualized using chemiluminescence reagent kit (Amersham Bioscience). The relative densities of the bands were quantified using ImageJ software (NIH).

CCl₄ was obtained from Sinopharm Chemical Reagent Co. Ltd. (Shanghai, China); APAP was obtained from Shanghai Aladdin Bio-Chem Technology Co. Ltd. (Shanghai, China); LaiPuRui (LPR) was obtained from Tonghua Renmin Pharmaceutical Co. Ltd. (Jilin, China); RPMI 1640 was purchased from Gibgo Company (Thermo Fisher Scientific, MA, USA); kits for detecting alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity, ROS, malondialdehyde (MDA), GSH, and SOD were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China); Trolox, DAPI, Triton-100, whole-cell protein extract, penicillin, and streptomycin were purchased from Solarbio Science & Technology Co. (Beijing, China); rhodamine 123 (Rh123), enhanced chemiluminescence kits were purchased from Beyotime Biotechnology (Shanghai, China); Alexa Fluor 488-congugated goat anti-rabbit IgG, Nrf2 rabbit antibody were obtained from Abcam (Cambridge, MA, USA); caspase-3 rabbit antibody, Bax rabbit antibody, and Bcl-2 rabbit antibody were all purchased from Cell Signaling Technology (Danvers, MA, USA); rabbit anti-β-action and anti-rabbit IgG were obtained from Proteintech Group, Inc. (Wuhan, China); Lipofectamine 2000 was bought from Invitrogen (Carlsbad, CA, USA); Nrf2 siRNA was purchased from Generaybiotech Co. Ltd. (Beijing, China). All other reagents used in this study were analytically pure.