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2-D Difference Gel Electrophoresis – an accurate quantitative method for protein analysis

Part 3. Proteomics

3.2. Expression Proteomics

Basic Techniques and Approaches

  1. Edward Hawkins,
  2. Stephen O. David

Published Online: 15 NOV 2005

DOI: 10.1002/047001153X.g302404

Encyclopedia of Genetics, Genomics, Proteomics and Bioinformatics

Encyclopedia of Genetics, Genomics, Proteomics and Bioinformatics

How to Cite

Hawkins, E. and David, S. O. 2005. 2-D Difference Gel Electrophoresis – an accurate quantitative method for protein analysis. Encyclopedia of Genetics, Genomics, Proteomics and Bioinformatics. 3:3.2:30.

Author Information

  1. GE Healthcare, Chalfont St. Giles, UK

Publication History

  1. Published Online: 15 NOV 2005

Abstract

2-D Difference Gel Electrophoresis (DIGE) is a novel technique that enables multiple protein extracts to be labeled with different fluorescent dyes. The labeled samples are then separated on the same 2-D gel. The dyes known as CyDyeTM DIGE Fluors are spectrally resolvable as well as size- and charge-matched.

The ability to add more than one sample to a 2-D electrophoresis (2-DE) gel enables the introduction of an internal standard that adds the quantitative advantages of a reference sample commonly seen in DNA microarray studies.

Measurement of protein abundance as a “Standardized Ratio” instead of an “Absolute volume” greatly reduces the influence of experimental variation commonly associated with conventional 2-DE.

Keywords:

  • 2-D DIGE;
  • DeCyderTM;
  • CyDyeTM DIGE Fluors;
  • TyphoonTM;
  • 2-D;
  • internal standard