Anti-RFP Antibody - Western Blot Western blot of RFP recombinant protein detected with polyclonal anti-RFP antibody. Lane 1 shows no reaction against a GFP recombinant protein present in 10 ug of HeLa cell extract. Lane 2 shows a single band detected in 10 ug of a HeLa lysate containing RFP recombinant protein. Separation was achieved using a 4-12% Bis-Tris gradient gel followed by transfer to nitrocellulose and blocking. The membrane was probed with the primary antibody diluted 1:2,500 for 1 h at room temperature followed by washes and reaction with a 1:5000 dilution of IRDye800 conjugated Goat-a-Rabbit IgG [H&L] MX (611-132-122). IRDye800 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.