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Updated: Aug 20, 2026

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid
Published on: August 14, 2009
Activity staining of glutathione peroxidase after two-dimensional gel electrophoresis
Chang Won Kho1, Sung Goo Park, Do Hee Lee
1Systemic Proteomics Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-600, Korea.
Abstract:
We have developed a method for rapid activity staining of proteins with glutathione peroxidase (GPx) activity after 2-D gel electrophoresis. After separating proteins extracted from yeast, or mouse red blood cells, by two-dimensional gel electrophoresis, SDS was removed and the gel was submerged in a Tris-HCl buffer containing glutathione and hydrogen peroxide, followed by incubation with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and phenazine methosulfate (PMS). After this proteins with GPx activity appeared as clear zones on a purple background. This relatively simple activity staining method could be useful for rapid screening of proteins with GPx activity in cell extracts.
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