Anti p src antibody

Tissue samples were collected from the ischemic hemisphere at 24 hours and 7 days, respectively, after reperfusion for the Western blot analysis. The samples at 24 hours after reperfusion were used for the detection of phosphorylated MAPK1 (p-ERK1/2) and phosphorylated RAC-alpha serine/threonine-protein kinase (p-AKT). The samples at 7 days after reperfusion were used for the detection of phosphorylated proto-oncogene tyrosine-protein kinase Src (p-SRC). Protein (40 μg) was electrophoresed on 10% SDS polyacrylamide gels (Beijing Biotides Biotechnology Co., Ltd, Beijing, China) and then transferred to a polyvinylidene fluoride membrane (Millipore Corporation, USA). The membrane was probed with primary antibody: anti-p-ERK1/2 antibody (Cell Signaling; 1 : 1000 dilution), anti-p-AKT antibody (Cell Signaling; 1 : 1000 dilution), and anti-p-SRC antibody (Cell Signaling; 1 : 1000 dilution). The specific reaction was visualized through the use of a chemiluminescent substrate (GE Healthcare, UK) [29 (link)]. β-Actin was used to verify equal loading. The optical density of protein was measured using ImageJ software (NIH, Bethesda, MD, USA) according to the manufacturer's instructions (n = 7 per group).