Activity staining of glutathione peroxidase after two-dimensional gel electrophoresis

Chang Won Kho, Sung Goo Park, Do Hee Lee, Sayeon Cho, Goo Taeg Oh, Seong Man Kang, Byoung Chul Park

Research output: Contribution to journalArticle

5 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)369-373
Number of pages5
JournalMolecules and Cells
Volume18
Issue number3
Publication statusPublished - 2004 Dec 31

Keywords

  • 2-D gel
  • Activity staining
  • Glutathione
  • Glutathione peroxidase

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

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  • Cite this

    Kho, C. W., Park, S. G., Lee, D. H., Cho, S., Oh, G. T., Kang, S. M., & Park, B. C. (2004). Activity staining of glutathione peroxidase after two-dimensional gel electrophoresis. Molecules and Cells, 18(3), 369-373.