Nitric Oxide Synthase Detection System, Fluorimetric
SIGMA/FCANOS1 - sufficient for 200 reactions
Product Type: Chemical
Induction of Nitric Oxide Synthase (NOS) activity in RAW 264.7 cells. RAW 264.7 cells were incubated for 20 hours with LPS and mouse g-interferon prior to the NOS activity measurement. The results shown are of the fluorescence detected after a 2 hour reaction. Excitation wavelength – 485 nm Emission wavelength – 530 nm RFU = Relative Fluorescence Units
| availability | not available in Japan |
| Quality Level | 200  |
| shipped in | wet ice |
| storage temp. | 2-8°C |
| usage | sufficient for 200 reactions |
| 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 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 2 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352303 |
| Components | Diaminofluorescein-2 diacetate (DAF2-DA) 1 mL/vial; Diphenyleneiodonium Chloride (DPI) 1 mM 200 μL; β-Nicotinamide adenine dinucleotide 2′-phosphate reduced tetrasodium salt, vial of 0.30-0.36 mg 1 vial; Arginine Substrate Solution for NOS Detection, Fluorimetric 2 mL |