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The current state of two‐dimensional electrophoresis with immobilized pH gradients
Author(s) -
Görg Angelika,
Obermaier Christian,
Boguth Günther,
Harder Alois,
Scheibe Burghardt,
Wildgruber Robert,
Weiss Walter
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(20000401)21:6<1037::aid-elps1037>3.0.co;2-v
Subject(s) - immobilized ph gradient , isoelectric focusing , chromatography , electrophoresis , chemistry , silver stain , resolution (logic) , analytical chemistry (journal) , computer science , microbiology and biotechnology , artificial intelligence , biology , biochemistry , enzyme
The original protocol of two‐dimensional electrophoresis with immobilized pH gradient (IPG‐Dalt; Görg et al. , Electrophoresis 1988, 9 , 531—546) is updated. Merits and limits of different methods for sample solubilization, sample application (by cup‐loading or in‐gel 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.