Western blot using Beta-galactosidase antibody. shows detection of a band at ~117 kDa (lane 1) corresponding to the protein present in partially purified preparations. Approximately 50 ng of protein was separated on a 4-20% Tris-Glycine gel by SDSPAGE and transferred onto nitrocellulose. After blocking the membrane was probed with the primary antibody diluted to 1/1,000. Reaction occurred overnight at 4°C followed by washes and reaction with a 1/10,000 dilution of IRDye800 (TM) conjugated goat-a-rabbit IgG [H&L] for 45 min at RT (800 nm channel, green). Molecular weight estimation was made by comparison to prestained MW markers in lane M (700 nm channel, red). RDye800 (TM) fluorescence image was captured using the Odyssey (R) infrared imaging system developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.
