Anti bax antibody sc 20067

Cells were plated in six-well plates. After treatment with ISL-17 (20 or 40 μM) and ISL (40 μM) for the indicated time, the cells were scraped from the plate. The collected protein samples were then electrophoresed on 10% SDS/PAGE gels, and electroblotted onto a PVDF membrane. At room temperature, the membranes were blocked with 5% nonfat milk for 2 h. Primary antibodies were incubated with membranes at 4°C overnight. Then, the membranes were washed with TBST for three times and incubated with secondary antibody at room temperature for 1 h. The immune-reactive complexes were analyzed with ECL kit (Bio-Rad, Hercules, CA, U.S.A.). The anti-Bcl-2 (sc-7382, 1:1000) and anti-Bax antibody (sc-20067, 1:1000) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, U.S.A.). Antibodies including anti-Cyclin B1 (#4135, 1:2000), anti-Cdc2 (#9116, 1:1000), anti-Cleaved PARP (#5625, 1:1000), anti-p62 (#8025, 1:1000), anti-LC3B (#3868, 1:1000), anti-Beclin 1 (#3495, 1:1000), anti-AKT (#4691, 1:1000), anti-phospho-AKT (#4060, 1:2000), anti-mTOR (#2983, 1:1000), anti-phospho-mTOR (#5536, 1:1000), anti-rabbit IgG-HRP (#7074, 1:1000) and anti-mouse IgG-HRP (#7076, 1:1000) were purchased from Cell Signaling Technology (Danvers, MA, U.S.A.).

Human proximal tubular epithelial cells (HK-2 cells) were cryopreserved at the Institute of Kidney Disease, Central South University. SS31 was synthesised and provided by Chinapeptide Co. Ltd. (Shanghai, China). Streptozocin (STZ) was obtained from Sigma-Aldrich (USA). The selective Drp1 inhibitor Mdivi1 (ab144589) was obtained from Abcam (UK). Anti-fibronectin (FN) antibody (sc-52331), anti-Bcl-2 antibody (sc-56015), anti-IL-1β antibody (sc-52012), and anti-Bax antibody (sc-20067) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-Drp1 rabbit monoclonal antibody (ab184247), anti-Mfn1 mice monoclonal antibody (ab57602), and Caspase1 antibody (ab138483) were purchased from Abcam (UK). The TUNEL assay kit (ab66110) and anti-β-actin antibody (ab8226) were purchased from Abcam (UK). Secondary antibodies in this study were purchased from KangChen Bio-tech (Shanghai, China). Other materials, including bovine serum albumin and low-glucose DMEM medium, were purchased from Gibco (USA).