| Assay Method Information | |
| | GLP-1R ACTIVATION - cAMP Assay 2 |
| Description: | GLP-1R activation by small molecule agonists is quantified by measuring cAMP increase in CHO cells stably expressing GLP-1R (MultiSpan product # C1267-1a). 50nL of agonists are pre-spotted in a 10 point dose response onto 384-well plate (Corning product # CL3826) using the Labcyte Echo System. The cells are harvested and plated in assay buffer (HBSS (Corning product # 21-023-CV) with 20mM Hepes (Gibco product # 15630-080) and 0.1% BSA (Rockland Immunochemicals product # BSA-1000)) or 100% Human Plasma (Innovative Research product #50-643-396) at 1,000 cells/well, 10μL/well, onto the pre-spotted plates. The cells are then incubated for 30 minutes at 370C, 5% CO2. cAMP concentration increase is then detected using Cisbio s cAMP Gs Dynamic Kit (product # 62AM4PEC) according to the manufacturer s protocol. The response is plotted against the log of the agonist concentration and fitted to a sigmoidal equation to determine the EC50. |
| Affinity data for this assay | |
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