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DAF-FM DA is a cell-permeable fluorescent probe for the detection of intracellular nitric oxide (NO). Once inside the cell, it is hydrolyzed by cytosolic esterases to a cell-impermeant DAF-FM. DAF-FM reacts with NO+ equivalents, such as nitric anhydride (N2O3), which are formed by autoxidation of NO. Under aerobic conditions, DAF-FM can rapidly and irreversibly trap NO to yield highly fluorescent triazolofluoresceins (DAF-Ts). DAF-FM DA is photostable and stable in pH 5.5 to 12. It can be used to quantitatively monitor agonist-evoked NO synthesis in cultured HUVECs. DAF-FM DA may also be used to quantify NO levels in induced pluripotent stem cell-derived derived cardiomyocytes (iPSC-CM).
It is recommended to prepare stock solutions in DMSO.
|Extinction Coefficient (ε)||84000 M-1cm-1|
|pH Sensitivity||Independent of pH when pH >5.8|
|Application||Confocal imaging, Flow cytometry|
|Storage||Store at -20°C|
The technical data provided above is for guidance only. For batch specific data refer to the Certificate of Analysis.
Tocris products are intended for laboratory research use only, unless stated otherwise.
References are publications that support the biological activity of the product.
Kojima et al (1999) Fluorescent Indicators for Imaging Nitric Oxide Production. Angew.Chem.Int.Ed.Engl. 38 3209 PMID: 10556905
Sheng et al (2005) DAF-FM (4-amino-5-methylamino-2',7'-difluorofluorescein) diacetate detects impairment of agonist-stimulated nitric oxide synthesis by elevated glucose in human vascular endothelial cells: reversal by vitamin C and L-sepiapterin. J.Pharmacol.Exp.Ther. 315 931 PMID: 16093274
Almeida et al (2021) Sensing nitric oxide in cells: historical technologies and future outlook. ACS.Sens. 6 1695 PMID: 33871990
Xu et al (2022) Uncompensated mitochondrial oxidative stress underlies heart failure in an iPSC-derived model of congenital heart disease. Cell Stem Cell 29 840 PMID: 35395180
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