Nitric Oxide Generation Kit

This kit contains 10 mg each of the following nitric oxide donors: SIN-1,

SNAP, S-nitrosoglutathione, spermine NONOate, and DEA-NONOate

Instruct ion Manual

Catalog Number PK-CA707-00239 Kit contains 5 x 10 mg (10 mg each of the following nitric oxide donors: Quantity SIN-1, SNAP, S-nitrosoglutathione, spermine NONOate, and DEA-NONOate)

Individual Kit Components Product Name SIN-1

Molecular Formula C6H10 N4O Molecular Weight 206.7 Molecular Structure

Synonyms Morpholinosydnonimine, hydrochloride Purity  95% as determined by TLC. Formulation White solid. Solubility Soluble in water, methanol or DMSO. Applications SIN-1 releases nitric oxide and forms superoxide spontaneously under physiological conditions and stimulates cyclic GMP production. It has been shown to be a potent vasodilator in vivo and in vitro and to inhibit smooth muscle cell mitogenesis and proliferation.

The relationship between NO generated from SIN-1 and intracellular calcium has been studied using fluorescent calcium indictors, which are also available from PromoCell. Storage & Stability Store desiccated at -20°C. References 1) Nature 364, 626(1993); 2) J. Pharmacol. Exp. Ther. 248, 762(1989); 3) Am J Physiol, L9 (1994); 4) Life Sci 54, 1449 (1994); 5) Neuron 12, 1235 (1994).

Product Name SNAP

Molecular Formula C7H12 N2O4S Molecular Weight 220 Molecular Structure

Synonyms S-nitroso-N-acetylpenicillamine Purity  98% as determined by TLC. Formulation Light green solid. Solubility Soluble in water or DMSO. 2 Instruction Manual

Applications SNAP releases nitric oxide and forms superoxide spontaneously under physiological conditions and stimulates cyclic GMP production. It has been shown to be a potent vasodilator in vivo and in vitro and to inhibit smooth muscle cell mitogenesis and proliferation.

The relationship between NO generated from SNAP and intracellular calcium has been studied using fluorescent calcium indictors, which are also available from PromoCell. The NO released from SNAP has also been reported to stimulate calcium-independent synaptic vesicle release, which can be detected with FM1-43 (also available from PromoCell). Storage & Stability Store desiccated at -20°C. References 1) Nishio, et al. Biochem. Biophys. Res. Comm. 221(1), 163(1996); 2) Gross, et al. Neuropharcology 33(7), 915(1994). 3) Am J Physiol, L9 (1994); 4) Life Sci 54, 1449 (1994); 5) Neuron 12, 1235 (1994).

Product Name S-Nitrosoglutathione

Molecular Formula C10 H16 N4O7S Molecular Weight 336 Molecular Structure

Synonyms N-(N-L--glutamyl-S-nitroso- L-cysteinyl)-glycine Purity  98% as determined by TLC. Formulation Hygroscopic pink solid. Specifications Absorption: 336 nm; Extinction Coefficient: 900 Solubility Soluble in water or DMSO. Storage & Stability Spontaneously decomposes in solution. Store desiccated at -20°C. References Biochemistry 34, 7177(1995); Br. J. Pharmacol. 107, 745(1992).

Product Name Spermine NONOate

Molecular Formula C10 H26 N6O2 Molecular Weight 262.4 Molecular Structure

Synonyms NA Purity  98% Formulation Crystalline solid. Specifications Absorption: 252 nm; Extinction Coefficient: 8500 Solubility Soluble in water or DMSO. Applications Spermine NONOate dissociates to the free amine and nitric oxide in a pH-dependent manner following first order kinetics. To initiate the release of nitric oxide, add a portion of the stock alkaline solution of Spermine NONOate to excess buffer of pH 7.0-7.4. The half- life of Spermine NONOate is 39 minutes at 37°C in 0.1 M phosphate buffer (pH 7.4). The decomposition of the NONOate is nearly instantaneous at pH 5.1. Storage & Stability Hygroscopic. Store desiccated at -20°C. For long term storage, keep sealed at -80°C, and it will be stable for at least one year. References 1) J. Med. Chem 34, 3242-3247 (1991); 2) Cancer Res. 53, 564-567 (1993); 3) Proc. Natl. Acad. Sci. USA 90, 98139817 (1993).

Instruction Manual 3

Product Name DEA-NONOate

Molecular Formula C4H10 N3O2  C4H12 N Molecular Weight 206.3 Molecular Structure

Synonyms (2-(N,N-diethylamino)-diazenolate-2-oxide, diethylammonium salt) Purity  98% as determined by HPLC. Formulation Off-white solid. Solubility Soluble in DMSO (2 mg/ml); 100% EtOH (25 mg/ml); water; PBS pH 7.2 (2 mg/ml). Applications Releases nitric oxide spontaneously under physiological conditions. Useful for reliable NO generation in solution. Storage & Stability Store desiccated under nitrogen at -20°C. References Keefer, et al. Methods in Enzymeology 268, 281(1996); Maragos, et al. J. Med. Chem. 34(11), 3242(1991).

FIRST AID All components above are potentially harmful, while toxicity is unknown. Avoid prolonged or repeated exposure. Avoid getting in eyes, on skin, or on clothing. Wash thoroughly after handling. If eye or skin contact occurs, wash affected areas with plenty of water for 15 minutes and seek medical advice. In case of inhaling or swallowing, move individual to fresh air and seek medical advice immediately. See MSDS for more information.

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