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

Additional information

Kit Size

0.125mg, 0.25mg

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.


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.

      Red wine consumption improves in vitro migration of endothelial progenitor cells in young, healthy individuals
    • 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
    • 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
    • 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;
    • Development of a fluorescent indicator for nitric oxide based on the fluoroscein chromophore: H. Kojima, et al.; Chem. Pharm. Bull. 46, 373 (1998)Abstract
    • Detection and imaging of nitric oxide with novel fluorescent indicators: diaminofluoresceins: H. Kojima, et al.; Anal. Chem. 70, 2446 (1998) Abstract
    • Part# 4006: DAF-2DA, 1 Vial (NOS200-1) (2-8C)
    • Part# 4006: DAF-2DA, 2 Vials (NOS200-2) (2-8C)
    • Formula: C24H18N2O7
    • MW: 446.4
    • Appearance: Liquid
    • Real time detection of NOS activity.
    • Manufactured: by Daiichi Pure Chemicals Co. Ltd. Japan