Fura-2, AM Ester Fluorescent Calcium Indicator

To study the roles of calcium, a key intracellular messenger in cells, it is essential to quantitatively monitor its concentration. The most widely used method of Ca2+ detection is the use of fluorescent Ca2+ indicators. Fura-2, AM ester is a ratiometric fluorescent dye that binds to free intracellular calcium, and is the preferred dye for ratiometric imaging microscopy. Fura-2, AM ester is membrane-permeant and thus can be loaded into cells via incubation. Fura-2, AM ester itself does not bind Ca2+, but it is readily hydrolyzed to Fura-2 by endogenous esterases once it enters cells. Fura-2 fluorescence can be detected using flow cytometry, fluorescence microscopy, or certain fluoresence microplate readers. Upon calcium binding, the fluorescent excitation maximum of the indicator undergoes a blue shift from 363 nm (Ca2+-free) to 335 nm (Ca2+-saturated), while the fluorescence emission maximum is relatively unchanged at ~510 nm. A major use of Fura-2 is in drug discovery, for measuring intracellular calcium mobilization after activation of G protein-coupled receptors (GPCRs) and calcium channels.

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