2D DIGE Analysis of Protein Extracts from Muscle Tissue - Difference Gel Electrophoresis (DIGE): Methods and Protocols - page 156

Internal
standard

Control Treated

Cy5 Cy2 Cy3

Mix samples

2D-electrophoresis

Image Acquisition and Analysis

Protein ID

Fig. 1. Scheme of a DIGE experiment, using three fl uors and a single gel.

Fig. 2. Scheme of the DIGE labeling reaction.

method is referred to as “minimal labeling.” This ensures that the
stoichiometry of protein to fl uor, results in only 1-2% of the total
number of lysine residues being labeled.
After labeling, protein samples are mixed and separated on the
same 2D gel. Different protein extracts labeled with different