2.11 | Western blot
Protein lysates from tissue and cell
sources were separated by SDS-PAGE, and transferred to PVDF membranes at
4 °C. The blots were washed with TBST (TBS + 0.1% Tween-20), blocked
with 7% skim milk in TBST for 1 h, and incubated with the specific
primary antibodies overnight at 4 °C on a shaking table. Next, the
membranes were incubated with HRP-conjugated anti-mouse or anti-rabbit
IgG secondary antibodies (1:5000) in 2% skim milk for 2 h with shaking
at room temperature. The immunoblots were visualised with an ECL kit
(Zen-Bio, Inc., China).