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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 NOS activity.
  • Applications - Flow Cytometry, Fluorescence Plate Reader, Fluorescence Microscope
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.

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.
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
  • Formulation: DAF-2DA is dissolved in DMSO at approx 5mM.
  • Manufactured: by Daiichi Pure Chemicals Co. Ltd. Japan

        Not For Sale in Japan

The following kits are available:
Catalog #
Size* Price
NOS200-1
0.125 mg per vial at 2.22 mg/mL. $125
NOS200-2 0.25 mg per vial at 2.22 mg/mL.
$245
* Please call 888 7 ASSAYS (888-727-7297) or email info@celltechnology.com for volume pricing
 
 Product Data Sheet
 Protocol
 MSDS