Language selection

Government of Canada / Gouvernement du Canada

Search

A modification of isocitrate and malate dehydrogenase assays for use in crude cell free extracts

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1016/0003-2697(73)90376-X
AuthorSearch for: 1; Search for: 1; Search for: 1
Affiliation
  1. National Research Council of Canada. Prairie Regional Laboratory
FormatText, Article
Abstract
A modification of the assays for isocitrate and malate dehydrogenase, using phenazine methosulphate and 2,6-dichlorophenolindophenol, permits measurements on cell-free extracts. Phenazine methosulfate at concentrations higher than 30 nmoles/3 ml prevents the accumulation of NADPH or NADH and thus reduces errors due to endogenous oxidation of these compounds. The use of 2,6-dichlorophenolindophenol rather than a tetrazolium salt as the terminal electron acceptor allows continuous spectrophotometric measurement of enzyme activities. Assay for NADP-specific isocitrate dehydrogenase can be performed in aerobic or anaerobic conditions. Assays for malate dehydrogenase should be run under anaerobic conditions because of the interference by oxygen on the phenazine methosulfate mediated reduction of 2,6-dichlorophenolindophenol by NADH. Under anaerobic conditions, where NADH oxidase is inoperative, the phenazine methosulfate/dichlorophenolindophenol assay is more sensitive than the assay using direct measurement of NADH at 340 nm.
Publication date
PublisherElsevier
In
  • Analytical Biochemistry 54, no. 2 (August 1973): 477–483.
LanguageEnglish
Peer reviewedYes
Identifier000326977390376X
NRCC-13170
Export citationExport as RIS
Report a correctionReport a correction (opens in a new tab)
Record identifiera99ec858-f63e-4f0a-a2af-0769bb664b2d
Record created2023-11-09
Record modified2023-11-09
Date modified: