Monoclonal caspase 3 antibody

Lysis of heart tissues was performed using the RIPA lysis buffer to extract proteins, followed by concentration measurement of the protein extracts and the BCA protein quantification kit was purchased from Thermo Fisher Scientific, Inc. (Waltham, MA, USA). A total of 5 µl of protein was loaded per lane. The proteins in the sample were separated using SDS-PAGE gel electrophoresis and transferred to the membrane (NC membrane; MilliporeSigma, Burlington, MA, USA) using a wet system. The primary antibodies [monoclonal Bax antibody (1:1,500; cat. no. YS-28034R) and monoclonal Bcl-2 antibody (1:1,000; cat. no. YSm-10846M) purchased from Santa Cruz Biotechnology, Inc., Dallas, TX, USA, and monoclonal caspase-3 antibody (1:500; cat. no. ab131000) purchased from Abcam, Cambridge, UK] were added after the membrane was blocked with 5% BSA, followed by incubation at 37°C overnight. Then, HRP-labeled goat anti-rabbit secondary antibody (1:1,000; cat. no. A0208, Beyotime Institute of Biotechnology, Shanghai, China) was added, and color was developed at 37°C. The percentages of separation and stacking gels were 10 and 5%, respectively.