Detection of Guaiacol Peroxidase on Electrophoretic Gels

Methods Mol Biol. 2017:1626:199-204. doi: 10.1007/978-1-4939-7111-4_18.

Abstract

It is possible to analyze peroxidase (POD) from different vegetable sources by electrophoresis. Zymography, i.e., a SDS-PAGE method to detect enzyme activity, is used to specifically detect POD activity and to visualize the total protein profile. For this purpose, we describe how a radish homogenate is prepared and submitted first to electrophoresis, and then, the POD activity present in the gel is reactivated and selectively stained using guaiacol as substrate. After scanning the gel, the same gel is further stained with Coomassie blue to determine the whole protein profile of the sample.

Keywords: Electrophoresis; Guaiacol peroxidase; Zymography.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Coloring Agents / analysis
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Enzyme Assays / methods*
  • Guaiacol / metabolism
  • Peroxidase / analysis*
  • Peroxidase / metabolism
  • Raphanus / enzymology*
  • Raphanus / metabolism
  • Rosaniline Dyes / analysis
  • Staining and Labeling / methods

Substances

  • Coloring Agents
  • Rosaniline Dyes
  • Guaiacol
  • Coomassie blue
  • guaiacol peroxidase
  • Peroxidase