Calcium Flux Assay Kit (Fura-2, No Wash, Ratiometric)

Product details

Calcium Flux Assay Kit (Fura-2, No Wash, Ratiometric) ab176766 is a no-wash, ratiometric calcium assay that allows homogeneous measurement of intracellular calcium mobilization caused by activation of G-protein-coupled receptors (GPCR) or calcium channels. The assay is suitable for use with 96-well and 384-well microplates.

In the Fura-2 calcium flux assay protocol, cells are pre-loaded with Fura-2 AM (which can cross the cell membrane). Once inside the cell, the lipophilic blocking groups of Fura-2 AM are cleaved by esterases, resulting in a negatively charged fluorescent dye that stays inside the cell.

The excitation maxima of the dye shifts from 363 nm to 335 nm upon binding of the dye to calcium. The emission spectra is relatively unaffected. The change is most easily measured by comparing the emission at 510 nm on excitation separately at 380 nm and 340 nm.

Measuring the ratio reduces the effects of uneven dye loading and cell numbers, dye leakage and photo bleaching, compared to measuring cellular calcium concentrations using a single wavelength dye like Fluo-8.

Fura-2 calcium flux assay protocol summary:
- add Fura-2 solution to cells
- incubate for 1 hr at 37°C, then for 20 min at room temp
- analyze with a microplate reader at both Ex/Em 340/510 nm and Ex/Em 380/510nm

This product is intended to be used for monitoring calcium fluctuations in vivo in live cells using the following HTS imaging plate readers: FLIPR™, FDSS, BMG NOVOstar™, FLexStation, ViewLux, IN Cell Analyzer or Arrayscan.