在聚丙烯酰胺凝胶中进行等电聚焦后，采用酶谱法检测弹性蛋白酶活性。
Detection of elastase activity with a zymogram method after isoelectric focusing in polyacrylamide gel.
作者信息
Gardi C, Lungarella G
出版信息
Anal Biochem. 1984 Aug 1;140(2):472-7. doi: 10.1016/0003-2697(84)90196-9.
A zymogram method for detecting elastase activity following isoelectric focusing in polyacrylamide gel is described. After enzyme activity has been visualized, the gel itself is available for protein staining and for analysis in sodium dodecyl sulfate-polyacrylamide gel electrophoresis in second dimension. The zymogram method is suitable for detecting microgram amounts of elastase and has one step only. It can be used with the purified enzyme as well as with crude extracts of tissue containing elastases showing activity toward succinyl-(Ala)3-p-nitroanilide. By this method a major component of elastase in both porcine and rat pancreas was detected. In addition, two forms of elastase with isoelectric points of 8.2 and 8.8, respectively, were identified in rat leukocyte extracts.
本文描述了一种用于检测聚丙烯酰胺凝胶等电聚焦后弹性蛋白酶活性的酶谱法。酶活性显现后，凝胶本身可用于蛋白质染色以及二维十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析。该酶谱法适用于检测微克量的弹性蛋白酶，且仅需一步操作。它可用于纯化的酶以及含有对琥珀酰-（丙氨酸）3-对硝基苯胺有活性的弹性蛋白酶的组织粗提物。通过这种方法，检测到了猪和大鼠胰腺中弹性蛋白酶的主要成分。此外，在大鼠白细胞提取物中鉴定出了两种等电点分别为8.2和8.8的弹性蛋白酶形式。
Zotero 插件