Faster and sensitive zymographic detection of oxidases generating hydrogen peroxide. The case of diamine oxidase

Anal Biochem. 2022 Jul 1:648:114676. doi: 10.1016/j.ab.2022.114676. Epub 2022 Mar 29.

Abstract

The existing zymography method for the detection of diamine oxidase (DAO) activity has been improved by a new staining procedure with the aim to ameliorate its sensitivity. Both procedures used SDS-PAGE gels containing uniformly distributed entrapped peroxidase (that wouldn't migrate during electrophoresis). The new approach with 3,5-dichloro-2-hydroxybenzenesulfonate (DCHBS) as peroxidase substrate and with 4-amino-antipyrine as color stabilizer allows a more sensitive detection of DAO when compared to the previously reported o-phenylenediamine (o-PDA) as peroxidase substrate. The newly improved method appears faster, simple and environmentally friendly. It can be used for most of oxidases releasing hydrogen peroxide as reaction product.

Keywords: Diamine oxidase; Dichloro-2-hydroxybenzenesulfonate (DCHBS); Glucose oxidase; Hydrogen peroxide; Peroxidase; Zymography.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amine Oxidase (Copper-Containing)*
  • Coloring Agents
  • Electrophoresis, Polyacrylamide Gel
  • Hydrogen Peroxide
  • Oxidoreductases
  • Peroxidase
  • Peroxidases

Substances

  • Coloring Agents
  • Hydrogen Peroxide
  • Oxidoreductases
  • Peroxidases
  • Peroxidase
  • Amine Oxidase (Copper-Containing)