The current state of two-dimensional electrophoresis with immobilized pH gradients
| Autor: | Angelika Görg, Günther Boguth, Robert Wildgruber, Burghardt Scheibe, Christian Obermaier, Walter Weiss, Alois Harder |
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| Rok vydání: | 2000 |
| Předmět: |
Silver Staining
Chromatography Two-dimensional gel electrophoresis Resolution (mass spectrometry) Chemistry Isoelectric focusing Clinical Biochemistry Analytical chemistry Proteins Sodium Dodecyl Sulfate Hydrogen-Ion Concentration Biochemistry Fluorescence Analytical Chemistry Electrophoresis Automation Solubilization Two dimensional electrophoresis Animals Electrophoresis Gel Two-Dimensional Immobilized pH gradient Gels Fluorescent Dyes |
| Zdroj: | Electrophoresis. 21(6) |
| ISSN: | 0173-0835 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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