| Assay Method Information | |
| | Biochemical Assay |
| Description: | hCB1/hCB1 b Gai Cyclic AMP (cAMP) Assays: Cells were seeded in a 384-well plate at an appropriate density from frozen cells. The cAMP assays were performed according to the manufacturer's protocol using MULTISCREEN™ TR-FRET cAMP 1.0 No Wash Assay Kit. Cells (hCB1 or hCB1 b) were pre-incubated with customer compounds for 30 minutes at rt followed by addition of EC80 CP55940 concentration and Forskolin. Cells were then incubated for 20 minutes at 37C. The reaction was terminated by sequentially adding sequentially adding MULTISCREEN™ Eu-labeled cAMP and MULTISCREEN™ 650-labeled anti-cAMP antibody in lysis buffer. The plate was then incubated at rt for 30 minutes before reading fluorescent emissions at 620 nm and 665 with excitation at 314 nm on FlexStation III (Molecular Devices). |
| Affinity data for this assay | |
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