Western blot analysis of Phospho-CBL (Y774) on different lysates with Rabbit anti-Phospho-CBL (Y774) antibody (HA722786) at 1/1,000 dilution. Lane 1: A549 cell lysate Lane 2: A549 starved for 2 hours cell lysate Lane 3: A549 starved for 2 hours then add 1μM sodium orthovanadate for 1 hour cell lysate Lane 4: HeLa cell lysate Lane 5: HeLa treated with 1mM sodium orthovanadate for 30 minutes cell lysate Lane 6: A549 starved for 2 hours then add 1μM sodium orthovanadate for 1 hour cell lysate, then the membrane treated with λpp for 1 hour Lane 7: HeLa treated with 1mM sodium orthovanadate for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 10 µg/Lane. Predicted band size: 100 kDa Observed band size: 100-110 kDa Exposure time: 59 seconds; ECL: K1802; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722786) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.