Figure 1. Multiplex detection of two target proteins by In-Gel Western. Two-fold dilutions of a T7-Tag extract (red; 700 nm) and purified Transferrin (green; 800 nm) were detected using the In-Gel Western protocol and IRDye® secondary antibodies. Gel was imaged with the Odyssey® CLx Imager. Reprinted from Urh, M. et al. Poster presentation. Advances in Genome Biology and Technology Conference (2002).
Primary and secondary antibodies must be carefully selected for two-color detection or cross-reactivity will result. Follow these guidelines for two-color detection:
Example: rabbit anti-protein X + mouse anti-protein Y primary antibodies
Example: goat anti-rabbit IgG + goat anti-mouse IgG
For two-color detection, follow Sample Protocol: One-color Detection with the following modifications: