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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

par Sener, Abdullah ;Malaisse, Willy
Référence Analytical biochemistry, 186, 2, page (236-242)
Publication Publié, 1990

Article révisé par les pairs

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Résumé :

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.