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Screen Quest™ Fluo-8 No Wash Calcium Assay Kit
Calcium flux assays are preferred methods in drug discovery for screening G protein coupled receptors (GPCR). Screen Quest™ Fluo-8 NW Calcium Assay Kit provides a homogeneous fluorescence-based assay for detecting the intracellular calcium mobilization. Cells expressing a GPCR of interest that signals through calcium are pre-loaded with our proprietary Fluo-8 NW which can cross cell membrane. Fluo-8 NW is the brightest calcium indicator available for HTS screening. Once inside the cell, the lipophilic blocking groups of Fluo-8 AM are cleaved by non-specific cell esterase, resulting in a negatively charged fluorescent dye that stays inside cells, and its fluorescence is greatly enhanced upon binding to calcium. When cells stimulated with screening compounds, the receptor signals release of intracellular calcium, which greatly increase the fluorescence of Fluo-8 NW. The characteristics of its long wavelength, high sensitivity, and 100-250 times fluorescence increases (when it forms complexes with calcium) make Fluo-8 NW an ideal indicator for measurement of cellular calcium. This Screen Quest Fluo-8 NW Calcium Assay Kit provides an optimized assay method for monitoring G-protein-coupled receptors (GPCRs) and calcium channels. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation.
Carbachol Dose Response was measured in HEK-293 cells with Screen Quest™ Fluo-8 No Wash Calcium Assay Kit and Fluo-4 NW Calcium Assay Kit. HEK-293 cells were seeded overnight at 40,000 cells/100 µL/well in a Costar black wall/clear bottom 96-well plate. The cells were incubated with 100 µL of dye-loading solution using the Screen Quest™ Fluo-8 No Wash calcium assay kit or Fluo-4 NW kit (according to the manufacturer's instructions) for 1 hour at room temperature. Carbachol (50µL/well) was added by NOVOstar (BMG Labtech) to achieve the final indicated concentrations.
Carbachol Dose Response was measured in HEK-293 cells with Screen Quest™ Fluo-8 No Wash Calcium Assay Kit and Fluo-4 NW Calcium Assay Kit. HEK-293 cells were seeded overnight at 40,000 cells/100 µL/well in a Costar black wall/clear bottom 96-well plate. The cells were incubated with 100 µL of dye-loading solution using the Screen Quest™ Fluo-8 No Wash calcium assay kit or Fluo-4 NW kit (according to the manufacturer's instructions) for 1 hour at room temperature. Carbachol (50µL/well) was added by NOVOstar (BMG Labtech) to achieve the final indicated concentrations.
protocol
Protocol
COA
CatalogSize
Price
Quantity
363141 Plate
Price
3631510 Plates
Price
36316100 Plates
Price
 
Spectral properties
Correction factor (260 nm)1.076
Correction factor (280 nm)0.769
Extinction coefficient (cm -1 M -1)
23430
Excitation (nm)495
Emission (nm)516
Quantum yield
0.16
1
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
Instrument settings
Fluorescence microplate reader
Excitation490 nm
Emission525 nm
Cutoff510 nm
Recommended plateBlack wall/Clear bottom
Instrument specification(s)Bottom read mode/Programmable liquid handling
Other instrumentsArrayScan, FDSS, FLIPR, FlexStation, IN Cell Analyzer, NOVOStar, ViewLux
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Page updated on October 13, 2025