title: The current state of two-dimensional electrophoresis with immobilized pH gradients.
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.
authors.affiliation: Proteomics Group, Institute of Food Technology and Analytical Chemistry, Technical University of Munich, Freising-Weihenstephan, Germany. angelika.gorg@lrz.tum.de
authors.forename: A
authors.initials: A
authors.lastname: Görg
authors.forename: C
authors.initials: C
authors.lastname: Obermaier
authors.forename: G
authors.initials: G
authors.lastname: Boguth
authors.forename: A
authors.initials: A
authors.lastname: Harder
authors.forename: B
authors.initials: B
authors.lastname: Scheibe
authors.forename: R
authors.initials: R
authors.lastname: Wildgruber
authors.forename: W
authors.initials: W
authors.lastname: Weiss
mesh_terms: D000818:Animals; D001331:Automation; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods* / Q000592:standards; D005456:Fluorescent Dyes; D005782:Gels; D006863:Hydrogen-Ion Concentration; D011506:Proteins / Q000032:analysis*; D016622:Silver Staining / Q000379:methods; D012967:Sodium Dodecyl Sulfate
chemical_list: D005456:Fluorescent Dyes; D005782:Gels; D011506:Proteins; D012967:Sodium Dodecyl Sulfate