DAF-FM (4-amino-5-methylamino-2′,7′-difluorofluorescein)
| Cat. # | Quantity | Price | Lead time | Buy this product |
|---|---|---|---|---|
| SB100 | 50 ug |
$22.00
|
in stock | |
| 1B100 | 1 mg |
$365.00
|
in stock |
DAF-FM (4-amino-5-methylamino-2′,7′-difluorofluorescein) is a cell-impermeant, fluorescent probe for detecting and quantifying low concentrations of nitric oxide (NO). DAF-FM does not need to be activated by cytosolic enzymes and is suitable to detect NO in extracellular matrix.
The fluorescence quantum yield of DAF-FM is ~0.005, but it increases about 160-fold to ~0.81 after reacting with NO and forming a fluorescent benzotriazole (excitation/emission maxima at 495/515 nm) .
The NO detection limit of DAF-FM (~3 nM) is more sensitive than that of DAF-2 (~5 nM). The fluorescence of the NO adduct of DAF-FM is independent of pH above pH 5.5. Moreover, the NO adduct of DAF-FM demonstrates a significantly enhanced photostability compared to that of DAF-2, ensuring reliable results and additional time for imaging.
DAF-FM should be dissolved in DMSO and then used to prepare a working solution. Buffers containing bovine serum albumin (BSA) or phenol red can affect the fluorescence and should be used cautiously.
The typical concentration of the dye for cell imaging is 1–10 µM. The dye concentration depends on cell type, as well as time and temperature of incubation, and should be defined experimentally.
The cell-permeant version of DAF FM — DAF-FM DA is also available.
Fluorescence emission spectra of DAF-FM, free and in solution containing nitric oxide (NO)
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General properties
| Appearance: | yellow to brown solid |
| Molecular weight: | 412.35 |
| CAS number: | 254109-20-1 |
| Molecular formula: | C21H14F2N2O5 |
| Solubility: | good in methanol, DMSO, DMF and water; limited in water; poor in acetonitrile and methylene chloride |
| Quality control: | NMR 1H and HPLC-MS (90+%) |
| Storage conditions: | 24 months after receival at -20°C in the dark. Transportation: at room temperature for up to 3 weeks. Desiccate. |
| MSDS: | Download |
| Product specifications |
Spectral properties
| Excitation/absorption maximum, nm: | 490 |
| Emission maximum, nm: | 516 (dye+NO) |
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