Cal-520™ PBX Calcium Assay Kit, Optimized for Probenecid-Sensitive Cell Lines

Kit Key Features

• Increased S/B Ratio: Significantly higher signal/background ratio than any other fluorescent Ca2+assays.
• Convenient: No wash and no probenecid needed. Formulated for probenecid-sensitive cell lines.
• Versatile Applications: Compatible with many cell lines and targets without ligand or target interference.

Cal-520™ PBX Calcium Assay Kit is optimized for probenecid-sensitive cell lines. Probenecid, a standard inhibitor of nonspecific anion transport, is not required for dye-loading with this kit. The assay involves only one dye addition step and does not require washing, allowing easy implementation in a high throughput environment. Cal-520™ AM is a new colorless fluorescent calcium-sensitive dye with a significantly improved signal to noise ratio and intracellular retention. Cells expressing a GPCR or calcium channel of interest that signals through calcium are pre-loaded with Cal-520™ AM which can cross cell membrane. Once inside the cell, the lipophilic blocking groups of Cal-520™ AM are cleaved by esterases, resulting in a negatively charged fluorescent dye that stays inside cells. Its fluorescence is greatly enhanced upon binding to calcium. When cells stimulated with agonists, the receptor signals the release of intracellular calcium, which significantly increase the fluorescence of Cal-520™. The characteristics of its long wavelength, high sensitivity, and >100 times fluorescence enhancement, make Cal-520™ AM an ideal indicator for the measurement of cellular calcium. The high S/B ratio and better intracellular retention make the Cal-520™ calcium assay a robust tool for evaluating GPCR and calcium channel targets as well as for screening their agonists and antagonists. Cal-520™ PBX Calcium Assay Kits provide an optimized assay method for monitoring the 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. The assay involves only one dye addition step and does not require washing, allowing easy implementation in a high throughput environment.

Figure1. ATP dose response comparison in CHO-K1 cells measured with Cal-520™ PBX, BD™ PBX, FLIPR Calcium 4 or Fluo-4 Direct™ Calcium Assay Kit. CHO-K1cells were seeded overnight in 60,000 cells per 100 μL per well in a 96-well black wall/clear bottom costar plate. The cells were incubated with 100 μL of the Cal-520™ PBX Calcium Assay Kit or 3 competitors kits (According to the Manufacturer’s instructions without adding probenecid) for 1.5 hours at 37°C. ATP (50μL/well) was added by FlexStation (Molecular Devices) to achieve the final indicated concentrations.

Cal-520™ PBX Calcium Assay Kit, Optimized for Probenecid-Sensitive Cell Lines

                36400    10 plates

                36401    100 plates

Online datasheet : www.interchim.eu/rapid_search.php?search="cal-520 pbx"&rad=0