Rabbit anti occludin 1

Equal amounts of protein were taken for per lane (20 µg) and separated by electrophoresis on 8-12% SDS-PAGE gels. Proteins were transferred to PVDF membranes by electrophoresis and sealed with 5% skim milk in Tris-buffered saline containing 0.1% Tween-20 (TBS-T) for 1.5 h at room temperature.
Thereafter, membranes were incubated with different primary antibodies at 4℃ overnight, including rabbit anti-Occludin-1 (1:1000, Invitrogen), mouse anti-Claudin-5 (1:1000, Invitrogen), and rat anti-ZO-1 (1:500, Santa Cruz Biotechnology), rabbit anti-MMP-9 (1:1000, Abcam), rabbit anti-SLC7A11 (1:1000, Abclone), rabbit anti-HO-1 (1:1000, Abclone), rabbit anti-GPX4 (1:1000, Abcam), rabbit anti-P2X7R (1:1000, Proteintech), rabbit anti-ERK1/2 (1:1000, Cell signaling technology), mouse anti-p-ERK1/2
(1:1000, Cell signaling technology), Rabbit anti-Albumin (1:1000, Proteintech), mouse anti-P53 (1:1000, Proteintech), rabbit anti-Transferrin (1:1000, Proteintech)and rabbit anti-GAPDH (1:3000, Abcam),. After washing with TBST for 3 times, PVDF membranes were immersed with horseradish peroxidase (HRP)conjugated secondary antibody (1:5000) for 1.5 h at room temperature. Immunolabeling were developed with enhanced ECL kit (Biosharp). Chemiluminescence levels was captured using an imaging system and data were normalized using GAPDH.