Key features and details
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Adherent cells, Suspension cells
Product nameFluo-4 Assay Kit (Calcium)
Sample typeAdherent cells, Suspension cells
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.
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.
Storage instructionsStore 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
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.
ab228555 has been referenced in 1 publication.
- Kim Y & Ghil S Regulators of G-protein signaling, RGS2 and RGS4, inhibit protease-activated receptor 4-mediated signaling by forming a complex with the receptor and Ga in live cells. Cell Commun Signal 18:86 (2020). PubMed: 32517689