Anti caspase3 ab32351

Proteins were extracted from lung tissues and quantified using the BCA protein concentration assay kit (Solarbio, China). Equal amounts of protein were resolved on sodium dodecyl sulphate-polyacrylamide gel electrophoresis gels, and transferred to polyvinylidene fluoride membranes. The membranes were blocked with 5% non-fat milk in TBST (10 mM Tris-HCl pH 7.4, 100 mM NaCl, 0.5% Tween-20) for 2 h at room temperature and incubated overnight at 4 °C with primary antibodies. The following primary antibodies were used according to the manufacturer’s protocol: anti-MMP9 (ab76003), anti-TIMP1 (ab109125), and anti-Caspase3 (ab32351) from Abcam (Cambridge, Cambridgeshire, UK); anti-Bax (#2772) from Cell Signaling Technology (Danvers, MA, USA); anti-GAPDH from Proteintech (Chicago, IL,USA); anti-MMP12 (bs-1854R) from Bioss (Beijing, China), Anti-Caspase8(WL02434), anti-Bcl2 (WL01556), and anti-Survivin (WL01684) from Wanleibio (Shenyang, Liaoning, China). The membranes were washed in TBST and incubated with secondary antibodies for 1.5 h. After extensive washing, the membranes were visualized using enhanced chemiluminescence reagent. Final images were analyzed using ImageJ software.