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

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

Paul Chomdom Kounga  1 , Armelle Tchoumi Neree  1 , Paola Pietrangeli  2 , Lucia Marcocci  2 , Mircea Alexandru Mateescu  3
Affiliations

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

Paul Chomdom Kounga et al. Anal Biochem. .

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.

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