Abstract
A sensitive and simple method for measuring NAD and NADH in the range of 0 to 200 μμmoles is described. The assay is based on the enzymic cycling of NAD and NADH and the sequential conversion of lactate-1- 14C to 14CO 2 in the presence of NAD or NADH, Mg ++, thiamine pyrophosphate, α-ketoglutarate, glutamic dehydrogenase, lactate dehydrogenase, and pyruvate dehydrogenase. Recovery of internal standard added to the tissue extract was approximately 100% and the assay of two different amounts of tissue resulted in equivalent concentrations of NAD or NADH per gram tissue.
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