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Fast horizontal electrophoresis. II. Development of fast automated staining procedures using PhastSystem™
Author(s) -
Olsson Ingmar,
Wheeler Richard,
Johansson Catherine,
Ekström Björn,
Stafström Nils,
Bikhabhai Rama,
Jacobson Gunilla
Publication year - 1988
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/elps.1150090105
Subject(s) - staining , silver stain , stain , silver nitrate , reagent , chemistry , chromatography , precipitation , fixation (population genetics) , nuclear chemistry , biochemistry , microbiology and biotechnology , biology , organic chemistry , genetics , physics , meteorology , gene
Abstract The development of equipment for fast automated staining is described. It is possible to handle staining procedures with up to 20 steps and nine different solutions. To increase the reaction rate in the reaction chamber, the gels are rotated and high temperatures are used. The temperature in the reaction chamber is controlled between room temperature and 50 °C. Increased temperature, above 20 °C, generally results in faster staining and destaining. However, some reactions proceed better at a low temperature, including fixation of proteins with TCA, and the development step in silver staining, where increased temperatures cause a high background stain. Silver in silver staining, where increased temperatures cause a high background stain. Silver staining using acidic silver nitrate solution is preferred, due to easy preparation and good storage stability of the reagents. This method also causes little precipitation of silver on the walls of the reaction chamber. Silver staining is accomplished within one hour. Staining with PhastGel Blue is accomplished within 30 min.