Fluo-3 AM is one of the most commonly used fluorescent probes for investigation of intracellular Ca²⁺ concentration. Its molecular formula is C₅₁H₅₀Cl₂N₂O₂₃ with a molecular weight of 1129.85.

Compared with Fura-2, Fluo-3 has several advantages. Firstly, it can be excited by a argon laser at 488nm, making it more convenient for detection. Secondly, Fluo-3 exhibits strong fluorescence change after binding to calcium, indicating that it is more sensitive to detect the change of Ca²⁺ concentration. Thirdly, Fluo-3 can detect higher concentration of calcium than Fura-2 due to its weaker affinity for calcium. Finally, its fluorescence change is more accurate for the immediate change of intracellular calcium, and the lag of fluorescence change caused by the slow dissociation from calcium is reduced.

This product is a 5mM stock solution formulated in anhydrous DMSO.

Fluo-3 AM is a cell-permeant dye, with very weak fluorescence that does not change with calcium concentration. After entering cells, Fluo-3 AM is cleaved by intracellular esterases to generate Fluo-3 which can be retained in cells. The calcium-binding Fluo-3 exhibits strong fluorescence with a maximum excitation wavelength of 506 nm and a maximum emission wavelength of 526 nm. Typically, we recommend using the excitation wavelength of 488nm and the emission wavelength of 525-530nm for the detection. Please refer to Figure 1 for the excitation and emission spectra of Fluo-3.
 

Figure 1. The excitation and emission spectra of Fluo-3

For the detection of intracellular calcium concentration, Fluo-3 AM is commonly used at concentrations of 0.5-5µM. Typically, fluorescent probe loading can be accomplished by incubating cells in 0.5-5µM of Fluo-3 AM for 15-60 min at 20-37℃, followed by appropriate washing, after which an additional 20-30min incubation can be performed to ensure sufficient conversion of Fluo-3 AM in cells.

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Store in the dark at -20℃ for up to 6 months.

This product will solidify and adhere to the interior wall of the vial at low temperatures, such as 4℃. Prior to use, it should be defrosted by incubation in a 20-25℃ water bath.

Please store this product away from light to minimize fluorescence quenching.

This product is for R&D only. Not for drug, household, or other uses.

For your safety and health, please wear a lab coat and disposable gloves during the operation.