一种用于飞克或纳克量胰凝乳蛋白酶的NADH偶联测定法。

An NADH-coupled assay for femtogram or nanogram quantities of chymotrypsin.

作者信息

Mroz E A, Lechene C

出版信息

Anal Biochem. 1983 Jan;128(1):181-5. doi: 10.1016/0003-2697(83)90360-3.

Abstract

Chymotrypsin can be determined with an NADH-coupled assay. Hydrolysis of the substrate benzoyltyrosine ethyl ester is monitored by coupling the liberation of ethanol to the production of NADH and determining the NADH spectrophotometrically or fluorometrically. Nanogram quantities of chymotrypsin can be determined in milliliter volumes. With these microfluorescence methods this assay can be performed in a final volume of less than a nanoliter, allowing determination of femtogram quantities of chymotrypsin, the amount present in an individual zymogen granule.

摘要

胰凝乳蛋白酶可用NADH偶联测定法进行测定。通过将乙醇的释放与NADH的产生相偶联，并采用分光光度法或荧光法测定NADH，来监测底物苯甲酰酪氨酸乙酯的水解。在毫升体积中可测定纳克量的胰凝乳蛋白酶。使用这些微量荧光方法，该测定可在最终体积小于一纳升的情况下进行，从而能够测定飞克量的胰凝乳蛋白酶，即单个酶原颗粒中的含量。