Standard Article

Gel Electrophoresis, 2D-difference

Proteins, Peptides and Amino Acids

  1. John F. Timms

Published Online: 15 SEP 2006

DOI: 10.1002/3527600906.mcb.200300094

Reviews in Cell Biology and Molecular Medicine

Reviews in Cell Biology and Molecular Medicine

How to Cite

Timms, J. F. 2006. Gel Electrophoresis, 2D-difference. Reviews in Cell Biology and Molecular Medicine. .

Author Information

  1. Ludwig Institute for Cancer Research, UCL Branch, London, UK

Publication History

  1. Published Online: 15 SEP 2006


Fluorescence two-dimensional difference gel electrophoresis (2D-DIGE) is a recently developed 2D-gel-based proteomics technique that can be applied to provide a sensitive, rapid, and quantitative analysis of differential protein expression between two or more biological samples. This technique utilizes chemical derivatives of spectrally distinct fluorescent cyanine dyes (Cy2, Cy3, and Cy5) to covalently label different samples prior to mixing and separation on the same 2-DE gels. This differential labeling and mixing of samples helps avoid the gel-to-gel variation inherent in conventional 2-DE-based expression profiling and reduces the number of gels that need to be run. Since the technique is also compatible with downstream identification of protein spots from gels by mass spectroscopy, it is particularly useful for reproducible, high-throughput proteomics.


  • Proteomics;
  • Two-dimensional Gel Electrophoresis (2-DE);
  • 2-DE Differential Protein Expression Analysis;
  • Fluorescence 2D-difference Gel Electrophoresis (2D-DIGE);
  • NHS-Cy2/Cy3/Cy5 Cyanine Dyes