NOS Detection Kit™
NOS (Nitric Oxide Synthase) Detection Kit

Key Benefits:
  • Cell permeable.
  • No wash homogenous assay.
  • Adaptable to High throughput assay platforms .
  • Real time detection of NO activity.
  • Applications - Flow Cytometry, Fluorescence Plate Reader, Fluorescence Microscopy
Assay Principle:

Diaminofluorescein –2 Diacetate (DAF-2DA) is a non-fluorescent cell permeable reagent that can measure free Nitric Oxide (NO) and nitric oxide synthase (NOS) activity in living cells under physiological conditions. Once inside the cell the diacetate groups on the DFA-2DA reagent arehydrolyzed by cytosolic esterases thus releasing FAD-2 and sequestering the reagent inside the cell. Production of nitric oxide converts the non-fluorescent dye, DAF-2, to its fluorescent triaole derivative, DAF-2T.

DAF-2T can be observed by: excitation 488nm and measuring emission at 515nm.
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Fig 1. DAF-2DA, the non-fluorescent probe, enters the cells where cytosolic esterases hydrolysis the diacetate groups, releasing the non-fluorescent DAF-2 dye. DAF-2 then reacts with NO and O2 to produce DAF-2T, its triazole fluorescent derivative.

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Fig 2. Phase Contrast and fluorescent microscopic images of rat aorta-derived vascular smooth muscular cells loaded with DAF-2DA .


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Product Data Sheet
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Protocol
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MSDS
Reactivity:

DAF-2DA can be used to detect NOS activity in cell culture or tissue sections. This reagent is not species specific and can also be used to detect NOS activity in plant cells.
Citations

Red wine consumption improves in vitro migration of endothelial progenitor cells in young, healthy individuals -Am J Clin Nutr 2010 92: 1 161-169; July 2010

References:
  1. Direct evidence of NO production in rat hippocampus and cortex using a new fluorescent indicator: DAF-2 DA: H. Kojima, et al.; Neuroreport 9, 3345 (1998) Abstract
  2. Direct evidence of nitric oxide production from bovine aortic endothelial cells using new fluorescence indicators: diaminofluoresceins: N. Nakatsubo, etal.; FEBS Lett. 427, 263 (1998) Abstract
  3. Nitric Oxide Ameliorates Hydrophobic Bile Acid-induced Apoptosis in Isolated Rat Hepatocytes by Non-mitochondrial Pathways: E. Gumpricht, et al.; J. Biol.Chem. 277, 25823 (2002) Full Text;
    http://www.jbc.org/cgi/content/full/277/28/25823#SEC1
  4. Development of a fluorescent indicator for nitric oxide based on the fluoroscein chromophore: H. Kojima, et al.; Chem. Pharm. Bull. 46, 373 (1998)Abstract
  5. Detection and imaging of nitric oxide with novel fluorescent indicators: diaminofluoresceins: H. Kojima, et al.; Anal. Chem. 70, 2446 (1998) Abstract
Product Specifications:
  • Formula: C24H18N2O7
  • MW: 446.4
  • Appearance: Liquid
  • Real time detection of NOS activity.
  • Manufactured: by Daiichi Pure Chemicals Co. Ltd. Japan

Not For Sale in Japan

 ProductCatalog #Size*Price
Nitric Oxide Synthase Detection KitNOS200-10.125 mg at 2.22mg/ml$145.00
Nitric Oxide Synthase Detection KitNOS200-2 0.25 mg at 2.22mg/ml$290.00

* Please call 888 7 ASSAYS (888-727-7297) or email info@celltechnology.com for volume pricing