FLUOFORTE® Calcium Assay Kits detect mobilization of intracellular calcium utilizing a fluorogenic calcium-binding dye optimized for superior cell-permeability and retention.
- Provides EC50 values comparable to Fluo-4 and Calcium 4
- Sensitive dye provides larger assay window allowing for detection of even weak signal compound responses
- Better able to detect native levels of GPCR expression in cultured cells
- Data suitable for comparison with that obtained using alternative dyes
The fluorogenic calcium-binding dye is provided to the cells as an acetoxymethyl (AM) ester, which is cell-permeable. Once inside the cells, the dye is hydrolyzed by intracellular esterases, which leads to generation of a cell membrane impermeable negatively charged form. In the absence of calcium, the calcium-binding moiety portion of the probe quenches the fluorescence of the fluorophore portion of the probe by photo-induced electron transfer. Binding of calcium relieves quenching and results in a several order of magnitude increase in the fluorescence emission intensity, with no shift in wavelength. The dye is capable of binding to physiologically relevant levels of calcium and increases in intracellular calcium lead to an increased fluorescence signal, which is readily measurable.
The self-quenching dye undergoes an electronic change upon binding of calcium, resulting in a several order of magnitude greater fluorescence.
Delivery information: Calcium Assay Kit (100x96 Tests) contains Reagent A (lyophilized FLUOFORTE® dye)(10 vials), Reagent B (dye efflux inhibitor) (10x10 mL).
Calcium Assay Kit (10x96 Tests) contains Reagent A (lyophilized FLUOFORTE® dye) (1 vial), Reagent B (dye efflux inhibitor) (10x1 mL), Reagent C (Hanks’ buffer with 20 mm HEPES).
