Endogenous Metabolite

NAD+是由连接腺苷5'-磷酸和核糖基烟酰胺5'-二磷酸的焦磷酸键组成的辅酶。 NADH 的氧化形式称为 NAD+ [1]。 NAD+ 在自然界中广泛存在，通过氧化 (NAD+) 和还原 (Nadide) 之间的交替，在众多酶活性中充当电子载体 [2]。

口服 NAD+ 补充剂已被用来治疗能量消耗、不明原因的疾病，如纤维肌痛和慢性疲劳综合症，以及简单的疲倦 [3]。

使用NADH和NAD+的氧化还原滴定。[3]
NADH在手套箱（O2<2ppm）中通过阴离子交换色谱法（5-ml HiTrap Q-Sepharose柱）重新纯化，以去除污染的NAD+。实验后，重新评估NADH储备溶液的完整性（在6小时内形成0.08±0.04%的NAD+）。通常，通过使用30μM NADH和不同量的NAD+（Sigma）来设置氧化还原电位，并通过使用NADH再生系统来检查低电位极限

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EPR。[3]
复合物I（10 mg ml−1）被1 mM纯化的NADH厌氧还原，或通过对纯化的NADH（≈−0.4 V）的透析还原，或使用1 mM NADH和10 mM NAD+还原至≈−0.3 V，并立即冷冻。使用ER 4119HS高灵敏度腔和ESR900连续流液氦低温恒温器[3]，在Bruker EMX X波段光谱仪上记录光谱。

NADH:ubiquinone oxidoreductase (complex I) is a major source of reactive oxygen species in mitochondria and a significant contributor to cellular oxidative stress. Here, we describe the kinetic and molecular mechanism of superoxide production by complex I isolated from bovine heart mitochondria and confirm that it produces predominantly superoxide, not hydrogen peroxide. Redox titrations and electron paramagnetic resonance spectroscopy exclude the iron-sulfur clusters and flavin radical as the source of superoxide, and, in the absence of a proton motive force, superoxide formation is not enhanced during turnover. Therefore, superoxide is formed by the transfer of one electron from fully reduced flavin to O2. The resulting flavin radical is unstable, so the remaining electron is probably redistributed to the iron-sulfur centers. The rate of superoxide production is determined by a bimolecular reaction between O2 and reduced flavin in an empty active site. The proportion of the flavin that is thus competent for reaction is set by a preequilibrium, determined by the dissociation constants of NADH and NAD+, and the reduction potentials of the flavin and NAD+. Consequently, the ratio and concentrations of NADH and NAD+ determine the rate of superoxide formation. This result clearly links our mechanism for the isolated enzyme to studies on intact mitochondria, in which superoxide production is enhanced when the NAD+ pool is reduced. Therefore, our mechanism forms a foundation for formulating causative connections between complex I defects and pathological effects.[3]

mouse LD50 intraperitoneal 4333 mg/kg Pharmaceutical Chemistry Journal, 20(160), 1986

[1]. Cellular and molecular mechanisms of metformin: an overview. Clin Sci (Lond), 2012. 122(6): p. 253-70.

[2]. Brandt, U., Energy converting NADH:quinone oxidoreductase (complex I). Annu Rev Biochem, 2006. 75: p. 69-92.

[3]. Kussmaul, L. and J. Hirst, The mechanism of superoxide production by NADH:ubiquinone oxidoreductase (complex I) from bovine heart mitochondria. Proc Natl Acad Sci U S A, 2006. 103(20): p. 7607-12.

NAD zwitterion is a NAD. It has a role as a geroprotector. It is functionally related to a deamido-NAD zwitterion. It is a conjugate base of a NAD(+).
A coenzyme composed of ribosylnicotinamide 5'-diphosphate coupled to adenosine 5'-phosphate by pyrophosphate linkage. It is found widely in nature and is involved in numerous enzymatic reactions in which it serves as an electron carrier by being alternately oxidized (NAD+) and reduced (NADH). (Dorland, 27th ed)
Nadide has been reported in Homo sapiens with data available.
Nadide is a dinucleotide of adenine and nicotinamide. It has coenzyme activity in redox reactions and also acts as a donor of ADP-ribose moieties.
A coenzyme composed of ribosylnicotinamide 5'-diphosphate coupled to adenosine 5'-phosphate by pyrophosphate linkage. It is found widely in nature and is involved in numerous enzymatic reactions in which it serves as an electron carrier by being alternately oxidized (NAD+) and reduced (NADH). (Dorland, 27th ed)

C21H27N7O14P2.3H2O

717.47

663.109

C, 38.02; H, 4.10; N, 14.78; O, 33.76; P, 9.34

53-84-9

NAD+-13C5 ammonium;NAD+-d4;NAD+-13C5-1;1859096-06-2;NAD+ lithium;64417-72-7; 53-84-9 (free acid); 20111-18-6 (sodium); 58-68-4 (reduced)

5892

White to off-white solid

140 - 142ºC

Gut microbial/endogenous metabolite

-5.72

340.71

7

18

11

44

1120

8

P(=O)(O[H])(OP(=O)([O-])OC([H])([H])[C@]1([H])[C@]([H])([C@]([H])([C@]([H])([N+]2=C([H])C([H])=C([H])C(C(N([H])[H])=O)=C2[H])O1)O[H])O[H])OC([H])([H])[C@]1([H])[C@]([H])([C@]([H])([C@]([H])(N2C([H])=NC3=C(N([H])[H])N=C([H])N=C23)O1)O[H])O[H]

BAWFJGJZGIEFAR-NNYOXOHSSA-N

InChI=1S/C21H27N7O14P2/c22-17-12-19(25-7-24-17)28(8-26-12)21-16(32)14(30)11(41-21)6-39-44(36,37)42-43(34,35)38-5-10-13(29)15(31)20(40-10)27-3-1-2-9(4-27)18(23)33/h1-4,7-8,10-11,13-16,20-21,29-32H,5-6H2,(H5-,22,23,24,25,33,34,35,36,37)/t10-,11-,13-,14-,15-,16-,20-,21-/m1/s1

1-((2R,3R,4S,5R)-5-((((((((2R,3S,4R,5R)-5-(6-amino-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)oxy)oxidophosphoryl)oxy)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-3-carbamoylpyridin-1-ium

β-DPNNSC-20272; Nadide; NSC 20272; NSC20272; beta-NAD; beta NAD; Enzopride Nadida; Codehydrase I; nadide; 53-84-9; NAD+; coenzyme I; beta-NAD; Codehydrogenase I; diphosphopyridine nucleotide; beta-nicotinamide adenine dinucleotide;NAD; NAD+; Nadidum; Nicotinamide; adenine dinucleotide

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ~41.67 mg/mL (~62.81 mM)

配方 1 中的溶解度: 100 mg/mL (150.73 mM) in PBS (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶。 (<60°C).

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。