The cAMP-Glo™ Assay is designed to monitor cAMP production in response to the effects of test compounds on G protein-coupled receptors (GPCR). GPCRs that couple with adenylate cyclase will increase or decrease intracellular cAMP. The assay is based on the principle that cyclic AMP (cAMP) stimulates protein kinase A (PKA) holoenzyme activity, decreasing available ATP and leading to decreased light production in a coupled luciferase reaction.

• Assay can be performed in 96-, 384-well plates and scalable to 1536-well plate formats and beyond
• Provides the best signal background ratio of all the cAMP assays
• Safer alternative for radioactive methods and no interference by fluorescent compounds

The cells are induced with a test compound to modulate cAMP levels and the cells are lysed to release cAMP, then the cAMP detection solution, which contains protein kinase A, is added. Kinase-Glo® Reagent is added to terminate the Protein kinase A reaction and detect the remaining ATP via a luciferase reaction. Plates are read using a microplate-reading luminometer. Luminescence can be correlated to cAMP concentrations using a cAMP standard curve. The half-life for the luminescent signal is greater than 4 hours. This extended signal half-life eliminates the need for luminometers with reagent injectors and allows batch-mode processing of multiple plates.