Dojindo,NOC 7/50/N377,CompoundsNOC 是稳定的 NO-胺复合物

Product Description of NOC CompoundsNOC 是稳定的 NO-胺复合物,可在生理条件下自发释放 NO,无需辅因子。 NO 释放速率取决于 NOC 的化学结构。与其他经典的 NO 供体(如硝酸甘油和 nitropurusside)相比,NOCs 自发生成 NO 的机制非常简单,并且副产物不会干扰细胞活动。一个 NOC 分子释放两个 NO 分子(如反应方案所示);第二个NO分子的释放速度很慢。 NOC 可用于以受控速率将受控数量的纯 NO 添加到实验系统中,且副作用最小。通过改变 NOC 试剂的浓度和选择,可以轻松控制释放的 NO 量。 Dojindo 提供具有不同半衰期的四种不同的 NOC(NOC 5、7、12 和 18)。在碱性溶液(如 NaOH 水溶液)中制备的 NOC 储备溶液相对稳定。然而,NOC 原液应在一天内使用,因为它每天降解约 5%,即使在 -20ºC 下也是如此。在将储备溶液添加到样品溶液后立即开始释放 NO。

Nitric Oxide Release1. 使用 0.1 M NaOH 制备 10 mM NOC 储备溶液。 由于NOC原液不稳定,所以放在冰浴上一天用完。 2. 将适量的 NOC 储备溶液添加到要释放 NO 的样品溶液中。 为保持样品溶液的 pH 值,NOC 储备溶液的体积不应超过样品体积的 1/50。 样品溶液应具有足够的缓冲作用。 加入 NOC 储备溶液后,NO 将立即释放。

Table 1 pH Dependency of NO Release at 37oC

1. K. Hayashi, et al., Action of Nitric Oxide as a Antioxidant Against Oxidation of Soybean Phosphatidylcholine Liposomal Membrane. FEBS Lett. 1995;370:37-40. (NOC 12)2. S. Shibuta, et al., Intracerebroventricular Administration of a Nitric Oxide-releasing Compound, NOC-18, Produces Thermal Hyperalgesia in Rats. Neurosci Lett. 1995;187:103-106. (NOC 18)3. S. Shibuta, et al., A new nitric oxide donor, NOC-18, exhibits a nociceptive effect in the rat formalin model. J Neurol Sci. 1996;141:1-5. (NOC 18)4. N. Yamanaka, et al., Nitric Oxide Released from Zwitterionic Polyamine/NO Adducts Inhibits Cu2+-induced Low Density Lipoprotein Oxidation. FEBS Lett. 1996;398:53-56. (NOC 5, NOC 7)5. D. Berendji, et al., Nitric Oxide Mediates Intracytoplasmic and Intranuclear Zinc Release. FEBS Lett. 1997;405:37-41.6. T. Ohnishi, et al., The Effect of Cu2+ on Rat Pulmonary Arterial Rings. Eur J Pharmacol. 1997;319:49-55. (NOC 7)7. Y. Adachi, et al., Renal Effect of a Nitric Oxide Donor, NOC 7, in Anethetized Rabbits. Eur J Pharmacol. 1997;324:223-226. (NOC 7)8. Y. Minamiyama, et al., Effect of Thiol Status on Nitric Oxide Metabolism in the Circulation. Arch Biochem Biophys. 1997;341:186-192. (NOC 7)
How do I prepare a stock solution?

Prepare 10-50 mM NOC solution with 0.1 M NaOH solution. Then add enough NOC solution to the cell culture to obtain a suitable concentration of NOC in cell culture. If the pH of the culture solution changes, use higher concentration of NOC.

What is the solubility of the NOC compounds?

NOC 5: 40 mg per 100 ml 0.1 M NaOH (2.2 M NOC 5)NOC 7: 70 mg per 100 ml 0.1 M NaOH (4.3 M NOC 7)NOC 12: 27 mg per 100 ml 0.1 M NaOH (1.5 M NOC 12)NOC 18: 20 mg per 100 ml 0.1 M NaOH (1.2 M NOC 18)

Is the stock solution stable?

The stock solution will lose 5% of its NOC activity per day, even when stored at -20oC. Please prepare fresh solution prior to use and keep the solution on an ice bath during the experiment. VI-1. Nitric Oxide Research: NO Donors

How is the half-life of NOC determined?

Prepare 20 mM NOC stock solution with 0.1 M NaOH. Warm PBS at 37ºC. Add 100 ml NOC solution to 1.9 ml PBS. Using a UV spectrophotometer, immediately start measuring its absorbance at the maximum wavelength of the NOC. Continue measuring until no further spectra changes are observed.

Can I use NOC for in vivo experiments?

Yes. Please review the papers by Shibata and colleagues (1995, 1996).

Is the amount of NO released in vitro the same as in vivo?

The amount of NO released in the solution should be the same if the pH and temperature are the same. However, the activity of NO may be different in vivo because of other reactive components such as thiol compounds and heme.