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DAF-FM (4-Amino-5-Methylamino-2',7'-Difluorofluorescein)

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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 cell-permeant version of DAF FM — DAF-FM DA is also available.

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6-Carboxy-H2DCFDA (6-carboxy-2',7'-dichlorodihydrofluorescein)

A reagent for detecting reactive oxygen species (ROS) in living cells. This carboxylated H2DCFDA analog has two additional negative charges that impede its leakage out of the cell.

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.
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