UNIT 10.23 Difference Gel Electrophoresis (DIGE) Using CyDye DIGE Fluor Minimal Dyes
Published Online: 1 FEB 2005
Copyright © 2005 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Chakravarti, B., Gallagher, S. R. and Chakravarti, D. N. 2005. Difference Gel Electrophoresis (DIGE) Using CyDye DIGE Fluor Minimal Dyes. Current Protocols in Molecular Biology. 69:VI:10.23:10.23.1–10.23.8.
- Published Online: 1 FEB 2005
- Published Print: JAN 2005
One- and two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (1- and 2-D SDS-PAGE) have been widely used for the separation and quantitative estimation of proteins. Following electrophoresis, the gels are stained appropriately to visualize the proteins. Difference gel electrophoresis (DIGE) is a new technique in which different protein samples, individually labeled with specific CyDyes, are combined together followed by electrophoresis and post electrophoretic co-detection and co-analysis on the same gel. CyDye DIGE fluor minimal dyes, which consist of three different CyDyes with different spectral characteristics, have been widely used for such purposes. The technique is highly sensitive with a wide dynamic range for detection of proteins and compatible with state-of-the-art protein identification techniques using mass spectrometry. Although DIGE is mainly used to compare differential expression of various protein samples using 2-D SDS-PAGE, 1-D DIGE also has important applications in quantitative proteomic studies.
- one-dimensional DIGE;
- CyeDye DIGE fluor minimal dyes