在使用5,5'-二硫代双(2-硝基苯甲酸)的谷胱甘肽还原酶测定中使用酶标仪。

Use of a microplate reader in an assay of glutathione reductase using 5,5'-dithiobis(2-nitrobenzoic acid).

作者信息

Cribb A E, Leeder J S, Spielberg S P

机构信息

Division of Clinical Pharmacology and Toxicology, Research Institute, Hospital for Sick Children, University of Toronto, Ontario, Canada.

出版信息

Anal Biochem. 1989 Nov 15;183(1):195-6. doi: 10.1016/0003-2697(89)90188-7.

DOI:10.1016/0003-2697(89)90188-7

PMID:2619044

Abstract

The use of 96-well microtiter plates and a programmable microplate reader to measure glutathione reductase in an assay based on reduction of 5,5'-dithiobis(2-nitrobenzoic acid) by GSH generated from an excess of GSSG is described. Samples are prepared in 96-well plates and absorbance at 415 nm with a reference wavelength of 595 is determined every 30 s for 3 min. The rate of increase in absorbance is directly proportional to the amount of glutathione reductase in the sample. Activity in an unknown sample is determined from a standard curve. The assay is rapid and allows many small samples to be analyzed in replicates of two or more at the same time.

摘要

描述了使用96孔微量滴定板和可编程酶标仪，基于过量氧化型谷胱甘肽（GSSG）生成的谷胱甘肽（GSH）还原5,5'-二硫代双（2-硝基苯甲酸）来测定谷胱甘肽还原酶的方法。将样品制备于96孔板中，在3分钟内每隔30秒测定415nm处的吸光度，参比波长为595nm。吸光度的增加速率与样品中谷胱甘肽还原酶的量成正比。未知样品的活性通过标准曲线来确定。该测定方法快速，可同时对许多小样品进行两个或更多重复分析。