Diaminofluorescein –2 Diacetate (DAF-2DA) is a
non-fluorescent cell permeable reagent that can measure free
Nitric Oxide (NO) and nitric oxide synthase (NOS) activity
in living cells under physiological conditions. Once inside
the cell the diacetate groups on the DFA-2DA reagent
arehydrolyzed by cytosolic esterases thus releasing FAD-2
and sequestering the reagent inside the cell. Production of
nitric oxide converts the non-fluorescent dye, DAF-2, to its
fluorescent triaole derivative, DAF-2T .
DAF-2T can be observed by: excitation 488nm and measuring
emission at 515nm.