Hsp60 rabbit mab

Preparation of protein lysates from the tissues and the western blot analysis was performed as previously described [24 (link)]. In short, 20 µg of protein was loaded per lane in each of the western blots and electrophoretically separated on 10% SDS–PAGE gels. All proteins were transferred at 4°C overnight at 20 V to a nitrocellulose membrane (Bio-Rad), blocked with 5% Blotting Grade Blocker (Bio-Rad) at room temperature for 60 min, then incubated with primary antibody at 4°C overnight. Both primary antibodies (SLC6A14 Rabbit pAb, # AP16835 Abclonal; HSP60 Rabbit mAb #12165 Cell Signaling) were diluted 1000-fold. After washing with TBST, the membrane was incubated for 1 h at room temperature with goat anti-rabbit secondary antibody (Bio-Rad) at a dilution of 3000-fold, washed in TBST again, then incubated for 5 min in Pierce ECL solution (ThermoFisher), before being visualized with autoradiography films.