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An enzymatic cycling method for nicotinamide-adenine dinucleotide with malic and alcohol dehydrogenases

✍ Scribed by Takahiko Kato; Sosamma J. Berger; Joyce A. Carter; Oliver H. Lowry

Publisher
Elsevier Science
Year
1973
Tongue
English
Weight
808 KB
Volume
53
Category
Article
ISSN
0003-2697

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✦ Synopsis

A method for measuring nicotinamide-adenine dinucleotide by enzymatic cycling is described which uses malic and alcohol dehydrogenases (EC 1.1.1.37, .and EC 1.1.1.1) for the enzyme couple. After cycling, malate is measured with either malic dehydrogenaze or malic enzyme (EC 1.1.1.40). The method has a number of advantages compared to those previously described. The cycling rate is high (greater than 30 OOO/hr) ; blank values are low; the reaction is linear over a wide range of NAD concentrations; and the terminal indicator reaction requires only one step. In addition the system is well suited for double cycling. This was shown by measurements of NAD in nuclei and cytoplasm from single dorsal root ganglion cells (rabbit). The overall amplification in this case was about 1 000 006.

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