Anti-GFP Antibody - Western Blot. Western blot of GFP recombinant protein detected with monoclonal anti-GFP antibody. GFP recombinant protein was expressed in HeLa cells, where 50 ng (lane 1), 100 ng (lane 2) and 500 ng (lane 3) of lysate were loaded per lane. Mab anti-GFP detects a 27 kD band corresponding to the epitope tag GFP. The cell lysates were prepared in a RIPA buffer containing 200 mM NaCl. A 4-12% Bis-Tris gradient gel (Invitrogen) was used for SDS-PAGE. The protein was transferred to nitrocellulose using standard methods. After blocking with 5% BLOTTO in PBS, the membrane was probed with the primary antibody diluted to 1.0 mg/ml for 1 h at room temperature followed by washes and reaction with a 1:2500 dilution of IRDye 800 conjugated Goat-a-Mouse IgG [H&L] MX10 (. 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.