Two-Color Western Blots
More Data, Improved Accuracy
The two infrared fluorescence detection channels of the Odyssey® Fc System enable two-color target analysis — an advantage that's not available with chemiluminescent or radioactive methods. Two-color Western analysis makes normalization easy and eliminates error introduced by stripping and reprobing or by comparison of separate blots. Superior image clarity and detail allow you to resolve co-migrating proteins and make it easier to detect subtle mobility shifts caused by protein modifications such as phosphorylation.
Figure 1. Detect Two Targets and Monitor Protein Phosphorylation. Lysates (10 µg/well) of A431 cells treated with EGF were separated and transferred to nitrocellulose. The blot was probed with rabbit anti-ERK1 and mouse anti-phospho-ERK primary antibodies (Santa Cruz Biotechnology) and then detected with goat anti-rabbit IRDye 680 (red) and goat anti-mouse IRDye 800CW (green) secondary antibodies, respectively. The blot was imaged with the Odyssey Fc Imager for 2 minutes in each channel. Overlapping ERK (red) and phospho-ERK (green) signals are displayed in yellow. This phospho-ERK1 antibody cross-reacts with phospho-EGFR (upper green band).