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

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

A Görg  1 , C Obermaier, G Boguth, A Harder, B Scheibe, R Wildgruber, W Weiss
Affiliations
Review

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

A Görg et al. Electrophoresis. 2000 Apr.

Abstract

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.

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