现有的用于检测二胺氧化酶(DAO)活性的酶谱方法已通过新的染色程序进行了改进,旨在提高其灵敏度。两种程序均使用含有均匀分布的包埋过氧化物酶(在电泳过程中不会迁移)的SDS-PAGE凝胶。与先前报道的邻苯二胺(o-PDA)相比,以3,5-二氯-2-羟基苯磺酸盐(DCHBS)作为过氧化物酶底物,以4-氨基-安替比林作为颜色稳定剂的新方法可以更灵敏地检测DAO作为过氧化物酶底物。新改进的方法看起来更快,简单环保。它可用于释放过氧化氢作为反应产物的大多数氧化酶。
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.