Objective: To develop nonradiometric, robust, and durable agonist assay for the activation of a target GPCR and a highly related GPCR.
Performance Specification: Dose-response with known agonist tool compound
Proposed Timeline: 3 months
Description: IBA took advantage of the fact that many activated GPCRs are coupled to the activation of phospho-MAPK1/MAPK2. The two target GPCR were stably expressed in cell culture and assessed for MAPK phosphorylation in response to known agonists.
Results: Successful expression and coupling of both GPCRs was demonstrated as assessed by western blotting cellular extracts with IBA custom made specific rabbit polyclonal antibodies and by showing a >10-fold increase in phospho-MAPK in response to known agonists.
Project Completion: 3 months