Castro Diana, Contreras Lellys M, Kurz Liliana, Wilkesman Jeff
Centre for Environmental, Biology and Chemistry Research, Facultad de Ciencias y Tecnología, Universidad de Carabobo, Valencia, Venezuela.
Methods Mol Biol. 2017;1626:199-204. doi: 10.1007/978-1-4939-7111-4_18.
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.
通过电泳分析不同蔬菜来源的过氧化物酶(POD)是可行的。酶谱法,即一种用于检测酶活性的SDS - PAGE方法,被用于特异性检测POD活性并可视化总蛋白谱。为此,我们描述了萝卜匀浆的制备方法,首先将其进行电泳,然后,凝胶中存在的POD活性被重新激活,并使用愈创木酚作为底物进行选择性染色。凝胶扫描后,同一凝胶再用考马斯亮蓝染色以确定样品的全蛋白谱。