Western blot analysis of mTOR on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (HA500126, 1/1,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: MCF-7 cell lysate Lane 2: Rat brain tissue lysate Lane 3: HepG2 cell lysate