Western blot analysis of HeLa lysate with anti-GAPDH antibody. Western blot analysis was performed on 20 μg HeLa lysate. Cells were lysed with RIPA buffer. The PVDF membrane was blocked with 5% dry milk in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-GAPDH antibody mouse monoclonal, clone 6C5 (Cat. No. 690975) was diluted in blocking buffer (antibody concentration 0.02 μg/ml) and incubated for 1 h at RT. The secondary antibody anti-mouse IgG goat polyclonal, HRP conjugate was also diluted in blocking buffer (antibody concentration 0.4 μg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using Pierce™ ECL Western Blotting Substrate.