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FCANOS1 Sigma

Nitric Oxide Synthase Detection System, Fluorimetric

sufficient for 200 reactions

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Description

Biochem/physiol Actions

Simple and specific assay for the measurement of free nitric oxide (NO) and nitric oxide synthase (NOS) activity in living cells under physiological conditions. Nitric oxide, formed from the amino acid L-arginine via the action of NOS, is a unique cell signaling molecule that functions as both an intracellular and an extracellular messenger. NO has been implicated in vasodilation, neurotransmission, cytotoxicity, and inflammation.
The standard assay for NOS activity is for solubilized cell-free enzyme preparations such as cell lysates, crude tissue extracts or purified enzymes. It measures the production of radio­labeled citrulline from radiolabeled arginine substrate (Sigma Product No. NOS-1). In contrast, the Fluorometric Cell-Associated NOS Detection System measures intracellular production of NO by a non-radiometric method. Radioisotopes are replaced by a cell-permeable diacetate derivative of 4,5-diaminofluorescein (DAF-2 DA). DAF-2 DA penetrates cells rapidly where it is hydrolyzed by intracellular esterase to DAF-2 that, in turn, reacts with NO produced by NOS to form a fluorescent triazolo-fluorescein. The fluorescent product can be quantitated using an excitation filter at 492 nm and an emission filter at 515 nm.

Legal Information

DAF-2 DA is manufactured by Daiichi Pure Chemicals Co., LTD.
US Patent No. 5,874, 590

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Flow Cytometry Instruments

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Diphenyleneiodonium Chloride (DPI) 1 mM 200 μL SDS    
Reaction Buffer 50 mL SDS    
Arginine Substrate Solution for NOS Detection, Fluorimetric 2 mL SDS    

Standard component

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Diaminofluorescein-2 diacetate (DAF2-DA) 1 mL/vial SDS D225
β-Nicotinamide adenine dinucleotide 2′-phosphate reduced tetrasodium salt, vial of 0.30-0.36 mg 1 vial SDS N9910
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
2
Protocols & Articles
Peer-Reviewed Papers
15

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