| Assay Method Information | |
| | hGLP-1R cAMP Assay |
| Description: | 1. Cell line and reagent preparation1) Cell line: Flp-In-293-GLP1R2) Medium: DMEM+10% FBS+1×Penicillin-Streptomycin+200 μg/mL HB3) Test buffer: 1×HBSS+20 mM HEPES+0.1% BSA+500 μM IBMX2. Agonist testa) Flpin-293-GLP1R cells were incubated in a 384-well assay plate (6007680-50, PE) using complete medium with 2,000 cells per well.b) Prepared a 4× complex working solution with assay buffer.c) Added 5 μL of 4× Compound Working Solution to the cell plate and incubated at 37° C. for 30 minutes.d) Diluted Eu-cAMP tracer (1/50) with lysis buffer and added 10 μl/well to the assay plate.e) Diluted Ulight-anti-cAMP (1/150) with lysis buffer and added 10 μl/well to the assay plate.f) Incubated at constant temperature for 1 hour.g) Read the plate on Envision 2105 plate reader at 665 nM and 615 nM. |
| Affinity data for this assay | |
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