Anti phosphorylated extracellular signal regulated kinase p erk 1 2

Samples from cells and animals were lysed in RIPA lysis buffer containing protease inhibitor and phosphatase inhibitor cocktail. The protein concentration was detected by BCA kits (Thermo Fisher Scientific, USA). Samples were separated by SDS-poly acrylamide gel electrophoresis and transferred to PVDF membrane. After blocked with 100 g/L non-fat milk for 2 h at room temperature, the membranes were incubated with primary antibody overnight at 4 °C, and then with secondary antibodies coupled to horseradish peroxidase. The primary antibodies were used for Western blot analysis:
anti-phosphorylated extracellular signal-regulated kinase (p-Erk) 1/2, anti-Erk1/2, antiphosphorylated p53, anti-p53 and anti-β-actin were purchased from Cell Signaling Technology (Boston, MA). Anti-Gpx4, anti-Bax, anti-Bcl-2 and anti-cleved caspase-3
were purchased from Abcam (UK). The secondary antibodies were used for Western blot analysis: anti-rabbit IgG-HRP and anti-mouse IgG-HRP (Santa Cruz Biotechnology, Dallas, TX, USA). Cross-reactivity was visualized using ECL Western blotting detection reagents and analyzed by scanning densitometry using a UVP Vision Works™ LS Software (UVP, Cambridge, UK) and quantified with ImageJ Software.