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Figure 2 | Proteome Science

Figure 2

From: Interactions by 2D Gel Electrophoresis Overlap (iGEO): a novel high fidelity approach to identify constituents of protein complexes

Figure 2

iGEO multi-color labeling scheme to identify PINCH-specific protein interactions. A, Tri-color iGEO labeling scheme to identify tag-independent PINCH binding partners in a single 2D gel. We applied CyDyes used for 2D-DIGE for protein labeling of Flag pull-down (red, Cy5), Protein A pull-down (green, Cy3), and Flag control (blue, Cy2). Spots containing both red and green hues, but devoid of blue hue are the spots of interest. B, A representative image of a 2D gel with the tri-color sample labeling. Twelve spots (circled in white) showed both red and green hues, but lacked blue hue. Arrows indicate bait proteins, PINCH-PrA (arrow 1), and Flag-PINCH (arrow 2). A source file of the gel image that allows inspection of signal intensity on each color channel is available as Additional file 4: Figure S1.

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Proteome Science

ISSN: 1477-5956