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

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

DAF-FM is a reagent that is used to detect and quantify low concentrations of nitric oxide (NO). It is essentiallyRead more
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D23844Promo Image10 x 50 μg
D23842Promo Image1 mg
Catalog number D23844
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10 x 50 μg
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Ask our AI about this Product
DAF-FM is a reagent that is used to detect and quantify low concentrations of nitric oxide (NO). It is essentially nonfluorescent until it reacts with NO to form a fluorescent benzotriazole. DAF-FM fluorescence can be detected by any instrument that can detect fluorescein, including flow cytometers, microscopes, fluorescent microplate readers and fluorometers.

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


Applications of DAF-FM:
• 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

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 ∼3 nM versus ∼5 nM for DAF-2).

Probes for Nitric Oxide Research
We offer several products for the detection of reactive oxygen species (ROS) and nitric oxide (NO). Review these products in Probes for Nitric Oxide Research—Section 18.3 in the Molecular Probes™ Handbook.

For Research Use Only. Not for human or animal therapeutic or diagnostic use.
For Research Use Only. Not for use in diagnostic procedures.
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Fluorescence spectra

Fluorescence spectra

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Lot #Certificate TypeDateCatalog Number(s)
3148259Certificate of AnalysisApr 09, 2025D23844
2916349Certificate of AnalysisAug 28, 2024D23842
2836703Certificate of AnalysisFeb 05, 2024D23844
2770843Certificate of AnalysisOct 04, 2023D23842
2550824Certificate of AnalysisMar 30, 2023D23842
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Citations & References (59)

Citations & References
Abstract
A stress surveillance system based on calcium and nitric oxide in marine diatoms.
Authors:Vardi A, Formiggini F, Casotti R, De Martino A, Ribalet F, Miralto A, Bowler C
Journal:PLoS Biol
PubMed ID:16475869
'Diatoms are an important group of eukaryotic phytoplankton, responsible for about 20% of global primary productivity. Study of the functional role of chemical signaling within phytoplankton assemblages is still in its infancy although recent reports in diatoms suggest the existence of chemical-based defense strategies. Here, we demonstrate how the accurate ... More
Nitric oxide-dependent mitochondrial biogenesis generates Ca2+ signaling profile of lupus T cells.
Authors:Nagy G, Barcza M, Gonchoroff N, Phillips PE, Perl A
Journal:J Immunol
PubMed ID:15356113
'Abnormal T cell activation and cell death underlie the pathology of systemic lupus erythematosus. Although mitochondrial hyperpolarization (MHP) represents an early and reversible checkpoint of T cell activation and apoptosis, lupus T cells exhibit persistent MHP. NO has recently been recognized as a key signal of mitochondrial biogenesis and mediator ... More
Fluorescence monitoring of ATP-stimulated, endothelium-derived nitric oxide production in channels of a poly(dimethylsiloxane)-based microfluidic device.
Authors:D'Amico Oblak T, Root P, Spence DM
Journal:Anal Chem
PubMed ID:16643013
'Intracellular nitric oxide (NO) production in a microfluidic endothelium is detected using fluorescence microscopy. Bovine pulmonary artery endothelial cells (bPAECs) were loaded with the fluorescence probe diaminodifluorofluorescein diacetate (DAF-FM DA), and the subsequent fluorescent DAF-FM DA/NO adduct was measured. Solutions of bradykinin, a well-known stimulus of endothelium-derived NO, activated nitric ... More
Carbohydrate oxidation acidifies endosomes, regulating antigen processing and TLR9 signaling.
Authors:Lewis CJ, Cobb BA,
Journal:J Immunol
PubMed ID:20200279
'Phagocytes kill encapsulated microbes through oxidative cleavage of surface carbohydrates, releasing glycan fragments and microbial contents that serve as ligands for immune receptors, which tailor the immune response against the offending pathogen. The glycan fragments serve as MHC class II (MHC II) ligands and innate receptor agonists, whereas microbial proteins ... More
Real-time measurement of nitric oxide in single mature mouse skeletal muscle fibres during contractions.
Authors:Pye D, Palomero J, Kabayo T, Jackson MJ
Journal:J Physiol
PubMed ID:17331997
'Nitric oxide (NO) is thought to play multiple roles in skeletal muscle including regulation of some adaptations to contractile activity, but appropriate methods for the analysis of intracellular NO activity are lacking. In this study we have examined the intracellular generation of NO in isolated single mature mouse skeletal muscle ... More
59 total citations

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