Counterion Dye Staining of Proteins in One- and Two-Dimensional Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis and Tryptic Gel Digestion of Stained Protein for Mass Spectrometry

  • Sun-Young Hwang
  • Jung-Kap ChoiEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1853)


A fast and matrix-assisted laser desorption/ionization-mass spectrometry compatible protein staining method in one- and two-dimensional sodium dodecyl sulfate–polyacrylamide gel electrophoresis is described. It is based on the counterion dye staining method that employs oppositely charged two dyes, zincon and ethyl violet 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 the sensitivity comparable to the colloidal Coomassie Brilliant Blue G stain using phosphoric acid as a component of staining solution (4–8 ng). The counterion dye stain does not induce protein modifications that complicate interpretation of peptide mapping data from mass spectrometry. Considering the speed, sensitivity, and compatibility with mass spectrometry, the counterion dye stain may be more practical than any other dye-based protein stains for routine proteomic researches.

Key words

Protein stain Proteomics 1-D and 2-DE Counterion dye stain MALDI-MS 



This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2016R1D1A3B03932150).


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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Laboratory of Analytical Biochemistry, College of Pharmacy & Research Institute of Drug DevelopmentChonnam National UniversityGwangjuSouth Korea

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