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A radiometric method for the determination of NADH in subpicomole amounts

Item Type:Article
Title:A radiometric method for the determination of NADH in subpicomole amounts
Creators Name:Weber, G. and Rosenthal, W. and Oberdisse, E.
Abstract:A radiometric method has been devised for the determination of small quantities of NADH formed in preceding dehydrogenase reactions. In a coupled enzymatic reaction, phosphoglycerate kinase (PGK) catalyzes the transfer of [32P]orthophosphate from [gamma-32P]ATP to 3-phosphoglycerate; the intermediate, 1,3-[1-32P]diphosphoglycerate, is dephosphorylated by glyceraldehyde-3-phosphate dehydrogenase (GAP-DH). [32P]Orthophosphate is released proportionally to NADH and can be measured after adsorption of [gamma-32P]ATP to activated charcoal. With this method, 0.2 pmol of NADH are detectable in the presence of a 10(4)-fold excess of NAD over NADH.
Keywords:NADH Determiation, ATP, [gamma-32P], Phosphoglycerate Kinase, Glyceraldehyde-3-Phosphate Dehydrogenase
Source:Journal of Biochemical and Biophysical Methods
ISSN:0165-022X
Publisher:Biophysical Society
Volume:15
Number:6
Page Range:295-306
Date:April 1988
Official Publication:https://doi.org/10.1016/0165-022X(88)90083-8
PubMed:View item in PubMed

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