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Screen Quest™ Luminometric Calcium Assay Kit
Calcium flux assays are preferred methods in drug discovery for screening G protein coupled receptors (GPCR). This kit uses a highly calcium-sensitive and membrane-permeable coelenterazine analog as a calcium indicator for the cells that are transfected with apoaequorin gene. Aequorin is a calcium-sensitive bioluminescent protein from the jellyfish Aequorea victoria that has been used extensively as a calcium indicator in cells. The aequorin complex emits blue light when bound to calcium ions. The luminescence intensity is directly proportional to the Ca2+ concentration. Our coelenterazine-based kit is much more sensitive than the fluorescence-based calcium assay kits (such as Fluo-4, Fluo-3, Calcium-3 and Calcium-4). This 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. It might be useful for monitoring of intracellular calcium mobilization in a specified compartment given that recombinant apoaequorin proteins can now be targeted to specific organelles, cells and tissues.
ATP Dose Response on CHO-aeq cells. CHO cells stably transfected with apoaequrin were seeded overnight at 50,000 cells/100 µL/well in a Costar white wall/clear bottom 96-well plate. The growth medium was removed and the cells were incubated with 100 µL of dye-loading solution using the Screen Quest™ Coelenterazine Calcium Assay Kit for 3 hours at room temperature and protected from light. ATP (25 µL/well) was added by NOVOstar (BMG Labtech) to achieve the final indicated concentrations. The EC50 of ATP is about 0.8 µM.
ATP Dose Response on CHO-aeq cells. CHO cells stably transfected with apoaequrin were seeded overnight at 50,000 cells/100 µL/well in a Costar white wall/clear bottom 96-well plate. The growth medium was removed and the cells were incubated with 100 µL of dye-loading solution using the Screen Quest™ Coelenterazine Calcium Assay Kit for 3 hours at room temperature and protected from light. ATP (25 µL/well) was added by NOVOstar (BMG Labtech) to achieve the final indicated concentrations. The EC50 of ATP is about 0.8 µM.
protocol
Protocol
CatalogSize
Price
Quantity
3630510 Plates
Price
36306100 Plates
Price
 
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
Instrument settings
Luminescence microplate reader
Recommended plateBlack wall/clear bottom
Instrument specification(s)Bottom read mode/Programmable liquid handling
Documents
Protocol
Safety Data Sheet (Catalog 36305)
Safety Data Sheet (Catalog 36306)
Certificate of Analysis
Brochure: Calcium Detection Probes & Assay Kits
Brochure: Physiological Probes & Assay Kits
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Page updated on October 30, 2025