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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.
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. €145
NOS200-2 0.25 mg per vial at 2.22 mg/mL.
 		 €290
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.
Fig 2. Phase Contrast and fluorescent microscopic images of rat aorta-derived vascular smooth muscular cells loaded with DAF-2DA .