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Fluo-4, AM ester(2mM)
Product overview:
Parameters
Appearance: Orange-red powder soluble in DMSO (F3013)
Ex/Em of Ca2+ –bound form: 494/516 nm
CAS No.: 273221-67-3
Molecular Formula: C51H50F2N2O23
Molecular Weight: 1096.94
Storage
Store at -20°C and protect from light. When stored as directed, product is stable for at least 12 months.
Description
Fluo-4, AM ester can enter into into cells via incubation. Fluo-4 AM ester itself does not bind Ca2+, but it is readily hydrolyzed to Fluo-4 by endogenous esterases once inside cells.
Fluo-4 is an analog of Fluo-3 with the two chlorine substituents replaced by fluorines, which results in increased fluorescence excitation at 488 nm that gives higher fluorescence signal. Fluo-4 has its absorption maximum at 494 nm, thus making it excitable by the argon-ion laser. Fluo-4 is essentially nonfluorescent without Ca2+ present, but the fluorescence increases at least 100 times on Ca2+ binding. Also, Fluo-4 was twice as strong as fluo-3 in fluorescence intensity. Fluo-4 AM ester can be used to detect intracellular free calcium by fluorescence microscopy, flow cytometry.
Notes
1. If a serum-containing medium is used, the esterase in the serum will decompose the AM , thereby reducing the effect of Fluo-4, AM entering the cells. In addition, the medium containing phenol red will slightly increase the background value, so you should try to remove the residue of the medium before adding the working solution.
2. There are quenching problems with fluorescent dyes. Please avoid light to slow down the fluorescence quenching.
3. This Fluo-4, AM is easy to absorb moisture. After removing it from the refrigerator, please make sure to open it in a dry environment at room temperature. As the reagent is extremely small, centrifuge it briefly before opening to ensure that the powder falls to the bottom of the tube.
4. Fluo-4, AM decomposes easily in contact with water. If it cannot be used up at one time, it is recommended to store the solution in small quantities.