| Assay Method Information | |
| | Functional Calcium Imaging Assay |
| Description: | Briefly, HEK 293T cells stably expressing the G protein-chimera Gα16gust44 were seeded onto 96-well plates coated with 10 μg/ml of poly-D-lysine. The next day cells were transiently transfected with ORA1, ORA2, or human bitter taste receptor TAS2R16 constructs using Lipofectamine 2000 (Invitrogen). After 24 h the cells were washed with buffer C1 (130 mM NaCl, 5 mM KCl, 10 mM Hepes, pH 7.4, 2 mM CaCl2, 10 mM glucose), and loaded with the calcium-responsive dye Fluo-4 AM in the presence of 1 mM probenecide, an inhibitor of ABC transporter A1. To remove excessive fluorescence dye, cells were washed three times with buffer C1 and transferred into a fluorometric imaging plate reader (FLIPR, Molecular Devices) for measurement. Test substances were dissolved in C1 buffer and the changes in fluorescence after application of test substances were monitored. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |