Journal
ANALYTICAL BIOCHEMISTRY
Volume 648, Issue -, Pages -Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2022.114676
Keywords
Diamine oxidase; Dichloro-2-hydroxybenzenesulfonate (DCHBS); Glucose oxidase; Hydrogen peroxide; Peroxidase; Zymography
Funding
- Fondation Courtois (Quebec, Canada)
- Natural Sciences and Engineering Research Council (NSERC) of Canada [06919]
- Ministere des Relations Internationales et de la Francophonie (MRIF, Quebec, Canada)
- Ministry of Foreign Affairs and International Cooperation (Italy)
- CRIPA/FRQNT (Fonds de recherche Nature et Technologies, Quebec, Canada)
- Fonds de la Recherche du Quebec-Sante (Quebec, Canada)
- CRIPA/FRQNT (Quebec, Canada)
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The existing zymography method for detecting DAO activity has been improved with a new staining procedure using 3,5-dichloro-2-hydroxybenzenesulfonate as peroxidase substrate and 4-amino-antipyrine as color stabilizer. This new method allows for a more sensitive detection of DAO compared to the previously reported substrate.
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 ophenylenediamine (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.
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