Immunoprecipitation analysis of 200ug extracts of A-431 cells, using 3 ug Phospho-EGFR-T669 pAb (AP0025). Western blot was performed from the immunoprecipitate using Phospho-EGFR-T669 pAb (AP0025) at a dilition of 1:1000. A-431 cells were treated by EGF (100 ng/mL) at 37℃ for 30 minutes after serum-starvation overnight.

Western blot analysis of extracts of A-431 and HUVEC cells, using Phospho-EGFR-T669 antibody (AP0025) at 1:1000 dilution. A431 cells were treated by EGF (100ng/mL) for 30 minutes after serum-starvation overnight. HUVEC cells were treated by EGF.|Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.|Lysates/proteins: 25ug per lane.|Blocking buffer: 3% BSA.|Detection: ECL Basic Kit (RM00020).|Exposure time: 10s.