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A sensitive radioisotopic method for the measurement of NAD(P)H: Its application to the assay of metabolites and enzymatic activities

Authors
Journal
Analytical Biochemistry
0003-2697
Publisher
Elsevier
Publication Date
Volume
186
Issue
2
Identifiers
DOI: 10.1016/0003-2697(90)90073-i

Abstract

Abstract A radioisotopic method for the assay of NADH or NADPH is presented, which is based on the conversion of 2-[U- 14C]ketoglutarate to 14C-labeled glutamate in the reaction catalyzed by glutamate dehydrogenase. The efficiency of the method is close to 75%, its precision (coefficient of variation) close to 5%, and its sensitivity close to 0.1 pmol/sample. This simple and rapid method can be applied to the measurement of several metabolites and enzymatic activities. In the present study, its application to the assay of sorbitol, 3-hydroxybutyrate, glutamate dehydrogenase, 3-hydroxybutyrate dehydrogenase, and glyceraldehyde-3-phosphate dehydrogenase is documented.

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