Western blot analysis of extracts of A-431 cells, using Phospho-SHC1-Y427 antibody (TA381474) at 1:1000 dilution.A-431 cells were treated by EGF (100 ng/ml) at 37‚ÑÉ for 30 minutes after serum-starvation overnight.|Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution.|Lysates/proteins: 25ug per lane.|Blocking buffer: 3% nonfat dry milk in TBST.|Detection: ECL Enhanced Kit .|Exposure time: 180s.