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Screen Quest™ Calbryte-590 Probenecid-Free and Wash-Free Calcium Assay Kit
Calcium flux assays are the preferred methods for screening G protein coupled receptors (GPCRs) in drug discovery. Screen Quest™ Calbryte-590™ Probenecid-Free and Wash-Free Calcium Assay Kit provides the most robust homogeneous red 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 Calbryte-590 AM which can cross cell membrane. Calbryte-590 AM is the brightest calcium indicator available for HTS screening. Once inside the cell, the lipophilic blocking groups of Calbryte-590 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 Calbryte-590 AM. The characteristics of its excellent cell retention, high sensitivity, and 100-250 times fluorescence increases (when it forms complexes with calcium) make Calbryte-590 AM an ideal indicator for measurement of cellular calcium. Calbryte-590 AM is the only red calcium dye that does not require probenecid to improve cellular retention. This Screen Quest™ Calbryte-590™ Probenecid-Free and Wash-Free Calcium Assay Kit provides the most optimized assay method for monitoring GPCRs and calcium channels with fragile or difficult cell lines. Compared to the green fluorescence-based calcium assays, this assay kit has less interference from the colored compounds from a compound library. It is also compatible with GFP cell lines for high content analysis. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation.
Graph illustrates signal-to-noise ratio (SNR) x 100%. ATP dose response was measured in CHO-K1 cells with Screen Quest™ Calbryte 590 Probenecid-Free and Wash-Free Calcium Assay Kit. CHO-K1 cells were seeded overnight at 50,000 cells/100 µL/well in a 96-well black wall/clear bottom costar plate. 100 µL dye loading solution was added and incubated for 45 min at 37°C and 15 min at RT. ATP (50 µL/well) was added by FlexStation 3 to achieve the final indicated concentrations.
Graph illustrates signal-to-noise ratio (SNR) x 100%. ATP dose response was measured in CHO-K1 cells with Screen Quest™ Calbryte 590 Probenecid-Free and Wash-Free Calcium Assay Kit. CHO-K1 cells were seeded overnight at 50,000 cells/100 µL/well in a 96-well black wall/clear bottom costar plate. 100 µL dye loading solution was added and incubated for 45 min at 37°C and 15 min at RT. ATP (50 µL/well) was added by FlexStation 3 to achieve the final indicated concentrations.
protocol
Protocol
CatalogSize
Price
Quantity
362001 Plate
Price
3620110 Plates
Price
36202100 Plates
Price
 
Spectral properties
Excitation (nm)581
Emission (nm)593
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
Excitation540 nm
Emission590 nm
Cutoff570 nm
Recommended plateBlack wall/clear bottom
Instrument specification(s)Bottom read mode/programmable liquid handling
Other instrumentsArrayScan, FDSS, FlexStation, IN Cell Analyzer, NOVOStar, ViewLux
Documents
Protocol
Safety Data Sheet (Catalog 36200)
Safety Data Sheet (Catalog 36201)
Safety Data Sheet (Catalog 36202)
Certificate of Analysis
Brochure: Calcium Detection Probes & Assay Kits
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Page updated on November 4, 2025