Using reporter gene technologies to detect changes in cAMP as a result of GPCR activation.
The measurement of cyclic adenosine monophosphate levels through reporter gene technology represents one of the most popular and cost-effective methods to assess changes in functional activity of G protein-coupled receptors (GPCRs). This chapter provides a generic protocol for the successful execution of a reporter gene assay for GPCRs, stably transfected within Chinese hamster ovary cell lines, signaling through the Gs pathway. It also highlights areas to investigate when developing reporter gene assays and additional factors to consider for assays that deviate from the protocol and cell line defined.
Rodrigues
Daniel J
DJ
Pfizer Global Research and Development, High Throughput Screening Centre of Emphasis, Sandwich, UK.
McLoughlin
David
D
D000818:Animals; D016466:CHO Cells / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D000242:Cyclic AMP / Q000032:analysis*; D017930:Genes, Reporter; D043562:Receptors, G-Protein-Coupled / Q000235:genetics*; D014162:Transfection; D001618:beta-Lactamases / Q000378:metabolism*
D043562:Receptors, G-Protein-Coupled; D000242:Cyclic AMP; D001618:beta-Lactamases