IndraLab

"Proteins were transferred onto PVDF (polyvinylidene difluoride) membranes in cold buffer (25 m m Tris Base, 192 m m glycine) by electrotransfer for 1.5 h, which were incubated in TBS buffer containing 5% milk for 1 h at 22–24 °C. The membrane was probed at 4 °C overnight with the following primary antibodies: Rabbit anti‐Mysm1 (1:1000, Abcam), rabbit anti‐PGC1 α (1:1000, Cell Signaling Technology), rabbit anti‐p‐p53 (1:1000, Cell Signaling Technology), rabbit anti‐p53 (1:1000, Cell Signaling Technology), rabbit anti‐p‐AMPK (1:1000, Cell Signaling Technology), rabbit anti‐AMPK (1:1000, Cell Signaling Technology), rabbit anti‐Sirt1 (1:1000, Cell Signaling Technology), rabbit anti‐p‐mTOR (1:1000, Cell Signaling Technology), rabbit anti‐mTOR(1:1000, Cell Signaling Technology), rabbit anti‐p‐S6 (1:1000, Cell Signaling Technology), mouse anti‐S6 (1:1000, Cell Signaling Technology), rabbit anti‐GAPDH (1:1000, Abclonal), mouse anti‐ β ‐actin (1:1000, APPLYGEN or Cell Signaling Technology)."