Sensitive assay for nicotinamide adenine dinucleotide phosphate and its reduced form based on the bioluminescence method |
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Authors: | José Ramacle Andrée Houbion Martine Raes |
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Affiliation: | Unité de Biochimie Cellulaire, Facultés Universitaires Notre-Dame de la Paix, B-5000 Namur Belgium |
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Abstract: | An assay of reduced nicotinamide adenine dinucleotide phosphate (NADPH) by bioluminescence was investigated and applied for NADP+. The NADP+ is first reduced by glucose-6-phosphate dehydrogenase and then assayed in a mixture containing a NADPH/flavin mononucleotide oxidoreductase which in turn activates luciferase. Many interferences were observed and the method was modified accordingly. NADP+ and NADPH can be assayed separately or simultaneously within the range 1–100 pmol, which is sufficiently sensitive to be applied to biological materials. Many details and precautions must be taken into consideration. |
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