Fluo-4 Assay Kit (Calcium) (ab228555)
Key features and details
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Adherent cells, Suspension cells
Overview
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Product name
Fluo-4 Assay Kit (Calcium) -
Detection method
Fluorescent -
Sample type
Adherent cells, Suspension cells -
Product overview
Fluo-4 Assay Kit (Calcium) (ab228555) provides a homogeneous fluorescence-based assay for detecting the intracellular calcium mobilization. The kit offers 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.
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Notes
Calcium flux assays are preferred methods in drug discovery for screening G protein coupled receptors (GPCR). Cells expressing a GPCR of interest that signals through calcium are pre-loaded with Fluo-4 AM which can cross cell membrane. Once inside the cell, the lipohilic blocking groups of Fluo-4 AM are cleaved by non-specific cell esterase, resulting in a negatively charged Fluo-4 dye that stays inside cells, and its fluorescence is greatly enhanced upon binding to calcium. When cells are stimulated with screening compounds, the receptor signals release of intracellular calcium, which greatly increase the fluorescence of Fluo-4.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 10 units 10X F127 Plus 1 x 10ml Fluo-4 AM 1 vial HHBS 1 x 100ml
Images
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ATP dose response was measured in CHO-K1 cells with Fluo-4 Assay Kit (Calcium)
CHO-K1 cells were seeded overnight at 60,000 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 Fluo-4 Assay Kit (Calcium) for 1 hour at 37oC, and then at room temperature for another 15 minutes. ATP (50µL/well) was added by Flexstation 3 to achieve the final indicated concentrations.