Differential Proteome Analysis Using 2D-DIGE

Methods Mol Biol. 2021;2228:77-84. doi: 10.1007/978-1-0716-1024-4_7.


Classical 2D-PAGE allows comparison and quantitation of proteomes by visualization of protein patterns using gel stains and comparative image analysis. The introduction of fluorescent reagents for protein labeling (difference in-gel electrophoresis or DIGE) has brought substantial improvement in this field. It provides multiplexing of up to three samples in one gel, higher sensitivity compared to normal protein staining methods, and a higher linear range for quantitation. This article gives detailed protocols for 2D-DIGE, including both minimal and saturation labeling.

Keywords: CyDye™; G-Dyes; Minimal labeling; S-Dyes; Saturation labeling; Two-dimensional difference in-gel electrophoresis (2D-DIGE); Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Electrophoresis, Polyacrylamide Gel*
  • Fluorescent Dyes
  • Humans
  • Luminescent Measurements
  • Proteins / analysis*
  • Proteome*
  • Proteomics*
  • Research Design
  • Staining and Labeling
  • Two-Dimensional Difference Gel Electrophoresis*


  • Fluorescent Dyes
  • Proteins
  • Proteome