The current state of two-dimensional electrophoresis with immobilized pH gradients

Electrophoresis. 2000 Apr;21(6):1037-53. doi: 10.1002/(SICI)1522-2683(20000401)21:6<1037::AID-ELPS1037>3.0.CO;2-V.


The original protocol of two-dimensional electrophoresis with immobilized pH gradient (IPG-Dalt; Gorg et al., Electrophoresis 1988, 9, 531-546) is updated. Merits and limits of different methods for sample solubilization, sample application (by cup-loading or ingel rehydration) with respect to the pH interval used for IPG-isoelectric focusing are critically discussed. Guidelines for running conditions of analytical and micropreparative IPG-Dalt, using wide IPGs up to pH 12 for overview patterns, or narrow IPGs for zoom-in gels for optimum resolution and detection of minor components, are stated. Results with extended separation distances as well as automated procedures are demonstrated, and a comparison between protein detection by silver staining and fluorescent dyes is given. A brief trouble shooting guide is also included.

Publication types

  • Review

MeSH terms

  • Animals
  • Automation
  • Electrophoresis, Gel, Two-Dimensional / methods*
  • Electrophoresis, Gel, Two-Dimensional / standards
  • Fluorescent Dyes
  • Gels
  • Hydrogen-Ion Concentration
  • Proteins / analysis*
  • Silver Staining / methods
  • Sodium Dodecyl Sulfate


  • Fluorescent Dyes
  • Gels
  • Proteins
  • Sodium Dodecyl Sulfate