Lane D A, Nadeau D
Département de biologie, Faculté des sciences, Université de Sherbrooke, Québec, Canada.
J Biochem Biophys Methods. 1988 Oct;17(2):107-17. doi: 10.1016/0165-022x(88)90041-3.
In this paper, we describe methods to assay specifically the NAD+, the NADP+, the NADH and the NADPH contents of cell monolayers. After an acid or an alkaline extraction, respectively for the oxidized or the reduced pyridine nucleotides, each type of nucleotide can be separately quantified with the use first, of specific reducing enzymes (lactate or glucose-6-phosphate dehydrogenases), followed by a bioluminescence enzymatic reaction [NADP(H)-FMN oxidoreductase and luciferase]. The assays hence developed are sensitive, reliable and specific. An application of the method with pulmonary alveolar macrophage monolayers is also described.
在本文中，我们描述了特异性测定细胞单层中NAD⁺、NADP⁺、NADH和NADPH含量的方法。分别对氧化型或还原型吡啶核苷酸进行酸提取或碱提取后，每种核苷酸均可先使用特异性还原酶（乳酸脱氢酶或葡萄糖-6-磷酸脱氢酶），然后通过生物发光酶促反应[NADP(H)-FMN氧化还原酶和荧光素酶]进行单独定量。由此开发的测定方法灵敏、可靠且特异。本文还描述了该方法在肺泡巨噬细胞单层中的应用。
