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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

Journal Article · · Analytical Biochemistry; (USA)
A radioisotopic method for the assay of NADH or NADPH is presented, which is based on the conversion of 2-(U-{sup 14}C)ketoglutarate to {sup 14}C-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.
OSTI ID:
6643958
Journal Information:
Analytical Biochemistry; (USA), Journal Name: Analytical Biochemistry; (USA) Vol. 186:2; ISSN ANBCA; ISSN 0003-2697
Country of Publication:
United States
Language:
English