DAF-FM is a reagent for the detection and quantification of low concentrations of nitric oxide (NO). DAF-FM itself does not fluoresce, but reacts with NO to form the fluorescent product benzotriazole. All instruments that can detect luciferin can detect DAF-FM fluorescence, including flow cytometers, microscopes, fluorescent microplate readers, and fluorometers.

DAF-FM quality standard:
•DAF-FM Ex/EM: ∼495/515 nm
• Lyophilized product should be dissolved in DMSO, then diluted with aqueous buffer, Made into working solution
• DAF-FM diacetate is cell-permeable and can passively penetrate cell membranes; once inside the cell, it converts to a cell-impermeable form
• Contains bovine serum white The buffer of protein (BSA) and phenol red may affect fluorescence and should be used with caution
• The fluorescence quantum yield of DAF-FM is ∼0.005, but after reaction with NO, the yield reaches about ∼0.81, which is an increase of about 160 times

Applications of DAF-FM:
• Assessment of NO production in transaldolase-deficient lymphoblastoid cells by flow cytometric analysis
• Detection of NO accumulation in embryonic cortical neurons after stimulation with neurotrophic factors
• In vivoimaging of NO in zebrafish
• Intravital microscopy to detect NO production associated with angiogenesis in mice
• Quantitative determination of ATP-induced NO release in rabbit platelets

Obvious advantages of DAF-FM over DAF-2
NO adduct spectrum and pH of DAF-FM Values above 5.5 are irrelevant. In addition, the photostability of the NO adduct of DAF-FM is significantly higher than that of DAF-2, which means that the image acquisition time needs to be extended. In summary, the DAF-FM reagent is more sensitive to NO than DAF-2 (for DAF-FM, the detection limit of NO is ∼3 nM; for DAF-2, the detection limit of NO is ∼5 nM).

Probes for Nitric Oxide Research
We offer a variety of products for detecting reactive oxygen species (ROS) and nitric oxide (NO). Available in the Molecular Probes™ Handbook Section 18.3—Probes for Nitric Oxide Detection.

For scientific research use only. Not for therapeutic or diagnostic use in humans or animals.