Unit

UNIT 10.23 Difference Gel Electrophoresis (DIGE) Using CyDye DIGE Fluor Minimal Dyes

  1. Bulbul Chakravarti1,
  2. Sean R. Gallagher2,
  3. Deb N. Chakravarti1

Published Online: 1 FEB 2005

DOI: 10.1002/0471142727.mb1023s69

Current Protocols in Molecular Biology

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.

Author Information

  1. 1

    Keck Graduate Institute of Applied Life Sciences, Claremont, California

  2. 2

    UVP, Inc., Upland, California

Publication History

  1. Published Online: 1 FEB 2005
  2. Published Print: JAN 2005

Abstract

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.

Keywords:

  • one-dimensional DIGE;
  • proteomics;
  • CyeDye DIGE fluor minimal dyes