Anti mouse igg 7076 secondary antibody

Western blotting was performed as previously described [47 (link)]. Primary antibodies for GCL-c (#390811), GCL-m (#55586), G6PDH (#373886), and β-actin (#47778) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-rabbit IgG (#2357) and anti-goat IgG (#2020) secondary antibodies were purchased from Santa Cruz Biotechnology, and anti-mouse IgG (#7076) secondary antibody was purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies were diluted in TBST (137 mM sodium chloride, 20 mM Tris, 0.1% Tween 20, pH 7.6.) containing 5% skim milk. Proteins in cell lysate samples were denatured by adding Laemmli 5× sample buffer and heating at 95 °C for 5 min. Proteins (40 μg) were resolved with 10% SDS-polyacrylamide gel electrophoresis at 80 V and electrically transferred to a polyvinylidene difluoride membrane (Amersham Pharmacia, Little Chalfont, UK) at 4 °C overnight. After blocking incubation with TBST containing 5% skim milk, the membrane was incubated with the primary antibody at 4 °C overnight, followed by incubation with the secondary antibody at room temperature for 1 h. The target protein bands were visualized with a chemiluminescence method using the picoEPD Western Reagent kit (ELPIS-Biotech, Daejeon, Korea). The captured blot images were analyzed using the Image J program from U.S. National Institutes of Health (Bethesda, MD, USA).