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

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.

• Ex/Em of DAF-FM: approx. 495/515nm
• 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 cell-impermeant for
• Buffers containing bovine serum albumin (BSA) and phenol red may affect fluorescence and should be used with caution
• Fluorescence quantum yield of DAF-FM is approx. 0.005, but increases about 160-fold, to ∼0.81, after reacting with NO
• 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
• Spectra of the NO adduct of DAF-FM are independent of pH above pH 5.5
• NO adduct of DAF-FM is significantly more photostable than that of DAF-2, which means additional time for image capture
• DAF-FM is a more sensitive reagent for NO than is DAF-2 (NO detection limit for DAF-FM approx. 3nM versus approx. 5nM for DAF-2)

Refer to the Molecular Probes™ Handbook for additional product information.

Cell Analysis, Cell Metabolism, Cell Viability, Proliferation and Function, Nitric Oxide Research, Nitro-Oxidative Stress