期刊
ELECTROPHORESIS
卷 25, 期 7-8, 页码 1136-1141出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/elps.200305776
关键词
counterion dye stain; matrix-assisted laser desorption/ionization mass spectrometry; one and two-dimensional gel electrophoresis protein stain; proteomics
A fast and matrix assisted laser desorption/ionization-mass spectrometry (MALDI-MS) compatible protein staining method in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1- and 2-D SDS-PAGE) is described. It is based on the counterion dye staining method that employs oppositely charged two dyes, zincon (ZC) and ethyl violet (EV) to form an ion-pair complex. The protocol, including fixing, staining and quick washing steps, can be completed in 1-1.5 h depending upon gel thickness. It has a sensitivity of 4-8 ng, comparable to that of colloidal Coomassie Brilliant Blue G (CBBG) staining with phosphoric acid in the staining solution. The counterion dye stain does not induce protein modifications that complicate interpretation of peptide mapping data from MS. Considering the speed, sensitivity and compatibility with MS, the counterion dye stain may be more practical than any other dye-based protein stains for routine proteomic researches.
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