Screen Quest(TM) Calbryte-590(TM) Probenecid-Free and Wash-Free Calcium Assay Kit

Ex: 573 nm. Em: 588 nm. Calcium flux assays are the preferred methods for screening G protein coupled receptors (GPCRs) in drug discovery. Screen QuestT Calbryte-590T Probenecid-Free and Wash-Free Calcium Assay Kit provides the most robust homogeneous red fluorescence-based assay for detecting the intracellular calcium mobilization. Cells expressing a GPCR of interest that signals through calcium are pre-loaded with our proprietary Calbryte-590 AM which can cross cell membrane. Calbryte-590 AM is the brightest calcium indicator available for HTS screening. Once inside the cell, the lipophilic blocking groups of Calbryte-590 AM are cleaved by non-specific cell esterase, resulting in a negatively charged fluorescent dye that stays inside cells, and its fluorescence is greatly enhanced upon binding to calcium. When cells stimulated with screening compounds, the receptor signals release of intracellular calcium, which greatly increase the fluorescence of Calbryte-590 AM. The characteristics of its excellent cell retention, high sensitivity, and 100-250 times fluorescence increases (when it forms complexes with calcium) make Calbryte-590 AM an ideal indicator for measurement of cellular calcium. Calbryte-590 AM is the only red calcium dye that does not require probenecid to improve cellular retention. This Screen QuestT Calbryte-590T Probenecid-Free and Wash-Free Calcium Assay Kit provides the most optimized assay method for monitoring GPCRs and calcium channels with fragile or difficult cell lines. Compared to the green fluorescence-based calcium assays, this assay kit has less interference from the colored compounds from a compound library. It is also compatible with GFP cell lines for high content analysis. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation.