Catalogue Number: 20651-AAT
|Shelf Life:||12 months|
Description: The intracellular calcium flux assay is a widely used method in monitoring signal transduction pathways and high throughput screening of G protein–coupled receptors (GPCRs) and calcium channel targets. Followed by Fluo-3 being introduced in 1989, Fluo-4, Fluo-8 and Cal-520 were later developed with improved signal/background ratio, and became the widely used Ca2+ indicators for confocal microscopy, flow cytometry and high throughput screening applications. However, there are still a few severe problems with Fluo-4. For example, as for Fluo-3, in all most all the intracellular calcium assays with Fluo-4 AM, probenecid is required to prevent the cell-loaded Fluo-4 from leaking out of cells. The use of probenecid with Fluo-4-based calcium assays compromises the assay results since probenecid is well-documented to have a variety of complicated cellular effects. Calbryte™ 520, AM is a new fluorescent and cell-permeable calcium indicator. Like other dye AM cell loading, Calbryte™ 520 AM ester is non-fluorescent and once gets inside cells, it is hydrolyzed by intracellular esterase and gets activated. The activated indicator is a polar molecule that is no longer capable of freely diffusing through cell membrane, essentially trapped inside cells. Upon binding Ca2+ ions, Calbryte™ 520 produces bright fluorescence signal with extremely high signal/background ratio. In addition, Calbryte™ 520 demonstrates greatly improved intracellular retention. It has the identical excitation and emission wavelength as Fluo-4, thus the same Fluo-4 assay settings can be readily applied to Calbryte™ 520-based calcium assays. Calbryte™ 520 is a new generation of fluorescent indicators for the measurement of intracellular calcium. Its greatly improved signal/background ratio and intracellular retention properties make Calbryte™ 520 AM the most robust indicator for evaluating GPCR and calcium channel targets as well as for screening their agonists and antagonists in live cells.