FLUOFORTE® Calcium assay kit

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Measurement of transient calcium mobilization from intracellular stores in response to the activation of G protein-coupled receptors (GPCRs) is considered a standard approach to the pharmacological characterization of receptors and compounds, frequently implemented in primary screening and lead development programs. The FLUOFORTE® Calcium Assay Kit is a member of the CELLESTIAL® product line, reagents and assay kits comprising fluorescent molecular probes that have been extensively benchmarked for live cell analysis applications.

A Brighter, More Robust Fluorescent Assay for Calcium Mobilization Dye optimized for superior cell-permeability and retention Economical alternative developed for use with conventional dual-dispensing microplate readers, i.e. BioTek, BMG Labtech, etcetera. Provides EC50 values comparable to Fluo-4 and Calcium 4 2-fold brighter fluorescence vs. Fluo-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   Fluo-Forte® Calcium Assay Kits detect mobilization of intracellular calcium utilizing a fluorogenic calcium-binding dye optimized for superior cell-permeability and retention. The self-quenching dye undergoes an electronic change upon binding of calcium, resulting in a several order of magnitude greater fluorescence. Mechanism of Action 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.

Flow Cytometry, Fluorescence microscopy, Fluorescent detection, HTS;Provides a homogeneous fluorescence-based assay for detecting intracellular calcium mobilization across a broad spectrum of biological targets