溶液的配制及保存.docVIP

NADH: Nicotinamide adenine dinucleotide (reduced form)中文名字：还原型烟酰胺腺嘌呤二核苷酸别名：Diphosphopyridine nucleotide，reduced form (DPNH)常见形式：Nicotinamide adenine dinucleotide, reduced dipotassium salt hydrate最大光吸收波长：340 nm、259 nm (pH 9.5) NAD（H）在细胞内如何执行功能呢？请参考以下文字介绍及Flash解说。 Nicotinamide adenine dinucleotide, abbreviated NAD+, is a coenzyme found in all living cells. The compound is a dinucleotide, since it consists of two nucleotides joined through their phosphate groups, with one nucleotide containing an adenine base and the other containing nicotinamide. In metabolism, NAD+ is involved in redox reactions, carrying electrons from one reaction to another. The coenzyme is therefore found in two forms in cells: NAD+ is an oxidizing agent – it accepts electrons from other molecules and becomes reduced. This reaction forms NADH, which can then be used as a reducing agent to donate electrons. These electron transfer reactions are the main function of NAD+. However, it is also used in other cellular processes, notably as a substrate of enzymes that add or remove chemical groups from proteins, in posttranslational modifications. Due to the importance of these functions, the enzymes involved in NAD+ metabolism are targets for drug discovery.?  NAD的结构式 NAD vs NADHNAD与NAD的最大光吸收波长比较 NAD溶液的配制及保存β-NAD可溶于水，浓度能够达到50 mg/ml，可能需微微加热。不过尽量不要加热，因为β-NAD遇热很容易降解，可用剧烈震荡或超声促进溶解。β-NAD溶液是酸性的，pH位于2～6之间，需分装后于-80 ℃冻存，每次用多少取多少，避免反复冻融。可以考虑加入NaOH或碳酸氢钠调pH至7左右，调节到中性的β-NAD水溶液在0 ℃可以保存2周左右。需要注意到是，β-NAD在碱性溶液中非常不稳定；遇到磷酸盐或碳酸盐也会变得不稳定；见光容易降解。另外，β-NAD极易受潮，以固体形态保存时务必要保持干燥。NADH溶液的配制及保存β-NADH可溶于0.01 M的NaOH溶液中，浓度可达100 mg/ml。不建议单独用水来配制β-NADH溶液，原因是β-NADH在水溶液中偏酸，容易降解。与β-NAD类似，β-NADH遇到磷酸盐也会变得很不稳定。β-NADH很容易被氧化，4 ℃使用时要注意以下两点：1、浓度不宜过高，≤5 mM 为适；2、pH 以9～11为宜。如果打算长期保存，浓度可以适当提高。建议：β-NADH应当现用现配，因为冻存时成分可能会有改变，而这些“杂质”在340 nm处的光吸收与β-NADH相同，且会抑制部分酶的活性。参考来源：Sigma公司的产品说明