All lanes : Anti-EGFR (phospho Y1086) antibody (ab227515) at 1/2000 dilution

Lane 1 : Serum starved (16 hours) A431 (human epidermoid carcinoma cell line) whole cell extract
Lane 2 : Serum starved (16 hours) A431 (human epidermoid carcinoma cell line) whole cell extract, treated with 100 ng/ml EGF for 5 minutes

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : HRP-conjugated anti-rabbit IgG

Developed using the ECL technique.

Predicted band size: 134 kDa

7.5% SDS-PAGE

4% paraformaldehyde-fixed A431 (human epidermoid carcinoma cell line) cells stained for EGFR (phospho Y1086) (green) using ab227515 at 1/500 dilution in ICC/IF.

Left-hand panels: A431 cells starved for 16 hours.

Right-hand panels: A431 cells starved for 16 hours then treated with 100 ng/ml EGF for 5 minutes.

Blue: Hoechst 33342 staining (lower panels).