| Assay Method Information | |
| | GLP-1R ACTIVATION - cAMP Assay 1 |
| Description: | GLP-1R activation by a compound of the present disclosure was quantified by measuring cAMP increase in CHO cells stably expressing GLP-1R (MultiSpan product # C1267-1a). The cells were harvested and plated in growth medium (DMEM/F-12 (Corning product # 10-090-CV) supplemented with 10% FBS (HyClone product # SH30071-03), penicillin/streptomycin (Corning product # 30-002CI) and 10μg/ml puromycin (Gibco product # A11138-03)) at 1,000 cells/well in a 384-well plate (Greiner product # 781080). The cells were then incubated overnight at 37 °C, 5% CO2. The next day, the medium was removed and the cells were washed with DPBS (Corning product # 21-031-CM) before adding the assay medium (HBSS, Corning product # 21-023-CV) with 20mM Hepes (Gibco product # 15630-080) and 0.1% BSA (Rockland Immunochemicals product # BSA-1000)). Following the medium change, the cells were incubated for 1 hour at 37 °C, 5% CO2. The tested GLP-1 compound was added to the cells in a 10 point dose response followed by a 30 minutes incubation at 37 °C, 5% CO2. cAMP concentration increase was then detected using Cisbio s cAMP Gs Dynamic Kit (product # 62AM4PEC) according to the manufacturer s protocol. The response was 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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