(4R)-Deuterated-reduced nicotinamide adenine dinucleotide phosphate, \((4 R)-\left[{ }^{2} H\right] N A D P H\), was prepared by reduction of NADP \({ }^{+}\)using an NADP \({ }^{+}\)-dependent alcohol dehydrogenase (EC 1.1 .1 .2 ) from Thermoanaerobium brockii and isopropanol- \(d_{8}\) as substrat
Preparation and storage of isotopically labeled reduced nicotinamide adenine dinucleotide
โ Scribed by Dexter B. Northrop; Ronald G. Duggleby
- Publisher
- Elsevier Science
- Year
- 1987
- Tongue
- English
- Weight
- 274 KB
- Volume
- 165
- Category
- Article
- ISSN
- 0003-2697
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โฆ Synopsis
A method for obtaining highly purified NADH in a dry, solid, and stable form is described. The method involves improvements of the ion-exchange and reversed-phase chromatographic procedures of C. J. Newton and S. M. Faynor, and D. B. Northrop (Anal. Biochem., 1983, 132, 50-53). The necessary time to prepare pure NADH has been reduced to a few hours. The final product, obtained by drying the nucleotide from absolute ethanol, shows no detectable decomposition either during the drying procedure or during storage under nitrogen gas at -20 degrees C for several months. Using dry product prepared from fixed volumes of ethanolic solution, standardized solutions of known amounts of the highly purified and stored NADH can be obtained in a few seconds.
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