Western Blot - Anti-GFP Antibody. Western blot of GFP recombinant protein detected with polyclonal anti-GFP antibody. Lane 1 shows detection of a 33 kD band corresponding to a GFP containing recombinant protein (arrowhead) expressed in HeLa cells. Lane 2 shows no staining of a mock transfected HeLa cell lysate. A 4-12% Bis-Tris gradient gel was used for SDS-PAGE. The protein was transferred to nitrocellulose using standard methods. After blocking the membrane was probed with the primary antibody diluted to 1 ug/ml for 1 h at room temperature followed by washes and reaction with a 1:2500 dilution of IRDye 800 conjugated Donkey-a-Goat IgG [H&L] MX7 (. The IRDye 800 fluorescence image was captured using the Odyssey Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.