Mittler R, Zilinskas B A
Department of Biochemistry and Microbiology, Cook College, Rutgers University, New Brunswick, New Jersey 08903-0231.
Anal Biochem. 1993 Aug 1;212(2):540-6. doi: 10.1006/abio.1993.1366.
A method for the detection of ascorbate peroxidase activity in native electrophoretic gels is described. The assay is based on the ability of ascorbate peroxidase to prevent the ascorbate-dependent reduction of nitroblue tetrazolium in the presence of H2O2. The method was found to be both sensitive (detection of less than 0.01 units of ascorbate peroxidase activity) and specific for ascorbate peroxidase activity. The application of the method for the detection of ascorbate peroxidase activity in protein extracts from several plant sources was investigated by comparing staining for activities of ascorbate peroxidase, horseradish peroxidase, and ascorbate oxidase and by immunodetection of ascorbate peroxidase in these extracts.
本文描述了一种在天然电泳凝胶中检测抗坏血酸过氧化物酶活性的方法。该测定基于抗坏血酸过氧化物酶在H2O2存在下防止抗坏血酸依赖性还原硝基蓝四唑的能力。该方法被发现既灵敏（可检测到低于0.01单位的抗坏血酸过氧化物酶活性）又对抗坏血酸过氧化物酶活性具有特异性。通过比较抗坏血酸过氧化物酶、辣根过氧化物酶和抗坏血酸氧化酶活性的染色以及对这些提取物中抗坏血酸过氧化物酶的免疫检测，研究了该方法在检测几种植物来源蛋白质提取物中抗坏血酸过氧化物酶活性方面的应用。
