Western blot using GeneTex's purified anti-p47 ING3 antibody shows detection of a band at ~55 kDa corresponding to ING3 in RKO cells transfected with ING3 (lane 2). Control RKO cells do not show detection of this specific band (lane 1). The identity of the non-specific bands at 33 kDa and 20 kDa has not been determined. Each lane contains approximately 10 μg of RKO whole cell lysate separated on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred to nitrocellulose. After blocking with 5% NF dry milk, the membrane was probed with the primary antibody diluted to 1:1,000. Incubation was at 4o C overnight followed by washes and reaction with a 1:20,000 dilution of IRDye?800 conjugated Rb-a-Goat IgG [H&L] MXHur for 45 min at room temperature. IRDye?800 fluorescence image was captured using the OdysseyR Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.