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Product information "DAF-FM DA / 3-Amino-4-(N-methylamino)-2',7'-difluorofluorescein diacetate "
3-Amino-4-(N-methylamino)-2',7'-difluorofluorescein diacetate. DAF-FM DA is an important reagent for quantification of low concentrations of nitric oxide in solution. This compound is essentially nonfluorescent until it reacts with NO to form a fluorescent benzotriazole. DAF-FM DA has an excitation/emission maxima of 495nm.
DAF-FM Specifications:
• Ex/Em of DAF-FM: ~495/515 nm
• Lyophilized product should be dissolved using DMSO and then added to an aqueous buffer to create a working solution.
• DAF-FM diacetate is cell permeant and passively diffuses across cellular membranes; once inside the cell, it is converted to a cell-impermeant form!
• Buffers containing bovine serum albumin (BSA) and phenol red may affect the fluorescence and should be used with caution.
• The fluorescence quantum yield of DAF-FM is ~0.005, but increases about 160-fold, to ~0.81, after reacting with NO.
Important Advantages of DAF-FM over DAF-2
The spectra of the NO adduct of DAF-FM are independent of pH above pH 5.5. Also, the NO adduct of DAF-FM is significantly more photostable than that of DAF-2 >, which means additional time for image capture. Finally, DAF-FM is a more sensitive reagent for NO than is DAF-2 (NO detection limit for DAF-FM ~3nM versus ~5nM for DAF-2).
Specifications:
Purity: >98% (HPLC)
Identity: determined by NMR
Appearance: light yellow powder
Fluorescence: λex 500 nm; λem 515 nm in DMSO
Soluble: in DMSO
C25H18F2N2O7
MW = 496.42 g/mol
Application:
• Assessment of NO production in transaldolase-deficient lymphoblasts by flow cytometry • Detection of NO accumulation in embryonic cortical neurons following neurotrophin stimulation • in vivo imaging of NO in zebrafish • Intravital microscopic detection of NO generation associated with angiogenesis in mice • Quantitation of ATP-induced NO release in rabbit platelets • Analyzation of NO production in mitochondriaSource:
syntheticClassification:
Documents:
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3-Amino-4-(N-methylamino)-2',7'-difluorofluorescein diacetate. DAF-FM DA is an important reagent for quantification of low concentrations of nitric oxide in solution. This compound is essentially nonfluorescent until it reacts with NO to form a fluorescent benzotriazole. DAF-FM DA has an excitation/emission maxima of 495nm. DAF-FM Specifications:• Ex/Em of DAF-FM: ~495/515 nm• Lyophilized product should be dissolved using DMSO and then added to an aqueous buffer to create a working solution. • DAF-FM diacetate is cell permeant and passively diffuses across cellular membranes; once inside the cell, it is converted to a cell-impermeant form! • Buffers containing bovine serum albumin (BSA) and phenol red may affect the fluorescence and should be used with caution.• The fluorescence quantum yield of DAF-FM is ~0.005, but increases about 160-fold, to ~0.81, after reacting with NO. Important Advantages of DAF-FM over DAF-2The spectra of the NO adduct of DAF-FM are independent of pH above pH 5.5. Also, the NO adduct of DAF-FM is significantly more photostable than that of DAF-2 >, which means additional time for image capture. Finally, DAF-FM is a more sensitive reagent for NO than is DAF-2 (NO detection limit for DAF-FM ~3nM versus ~5nM for DAF-2).
4,5-Diamino-N,N,N',N'-tetraethyl-rhodamine. Sensitive NO probe, LOD of 10 nM, shows higher photostability than the classical fluorescein derivative DAF.
4,5-Diaminofluorescein triazole (DAF-2T) can be used as a reference material for DAF-2 (S5439 >).References: (1) M. Feelisch et al;, Free Radical Biol. Med. 38(3), 356 (2005); (2) T. Nagano et al.; J. Biol. Chem. 277(1), 47 (2002); (3) T. Nagano et al.; FEBS Letters 427(2), 263 (1998); (4) H. Kojima et al.; Chem. Pharm. Bull. 46(2), 373 (1998)