WB
Western blot analysis of Phospho-PLC-gamma-1 (Y783) on different lysates with Rabbit anti-Phospho-PLC-gamma-1 (Y783) antibody at 1/1,000 dilution.
Lane 1: A431 cell lysate,
Lane 2: A431 starved overnight then add 20ng/mL EGF for 5 minutes cell lysate,
Lane 3: NIH/3T3 cell lysate,
Lane 4: NIH/3T3 starved overnight then add 100ng/mL PDGF for 5 minutes cell lysate,
Lane 5: A431 starved overnight then add 20ng/mL EGF for 5 minutes cell lysate, then the membrane treated with λpp for 1 hour,
Lysates/proteins at 20 µg/Lane.
Exposure time: 3 minutes;
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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.