Extraction and Measurement of NAD(P)+ and NAD(P)H
Extraction and Measurement of NAD(P)+ and NAD(P)H
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DOI：
10.1007/978-1-4939-0473-0_26
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发表时间：
2014-01-01
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影响因子：
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通讯作者：
Newman, Dianne K.
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文献类型：
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作者：
Kern, Suzanne E.;Price-Whelan, Alexa;Newman, Dianne K.
烟酰胺腺嘌呤二核苷酸是无数分解代谢和合成代谢反应的关键氧化还原活性底物。因此，NAD(+)/NADH和NADP(+)/NADPH的比值被认为是整体细胞内氧化还原电位和代谢状态的关键指标。这些比率可以在包括酸和碱溶液的简单提取程序之后，在散装条件下使用基于酶循环的高灵敏比色分析(检测下限为或低于0.05微米或1 pmol)来测量。有必要特别考虑避免因内源性氧化还原活性代谢物的存在而造成的测量伪影，例如由不同假单胞菌产生的吩嗪类物质(见第25章)。
Nicotinamide adenine dinucleotides are critical redox-active substrates for countless catabolic and anabolic reactions. Ratios of NAD(+) to NADH and NADP(+) to NADPH are therefore considered key indicators of the overall intracellular redox potential and metabolic state. These ratios can be measured in bulk conditions using a highly sensitive enzyme cycling-based colorimetric assay (detection limit at or below 0.05 mu M or 1 pmol) following a simple extraction procedure involving solutions of acid and base. Special considerations are necessary to avoid measurement artifacts caused by the presence of endogenous redox-active metabolites, such as phenazines made by diverse Pseudomonas species (see Chapter 25).