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Fast visible dye staining of proteins in one‐ and two‐dimensional sodium dodecyl sulfate‐polyacrylamide gels compatible with matrix assisted laser desorption/ionization‐mass spectrometry
Author(s) -
Choi JungKap,
Chae HoZoon,
Hwang SunYoung,
Choi HoonIn,
Jin LiTai,
Yoo GyurngSoo
Publication year - 2004
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.200305776
Subject(s) - staining , chemistry , coomassie brilliant blue , chromatography , counterion , mass spectrometry , stain , sodium dodecyl sulfate , polyacrylamide , matrix assisted laser desorption/ionization , polyacrylamide gel electrophoresis , sodium , desorption , analytical chemistry (journal) , ion , biochemistry , adsorption , polymer chemistry , organic chemistry , medicine , pathology , enzyme
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.