cAMP-Glo Assay

cAMP-Glo Assay is a homogeneous, bioluminescent, high-throughput assay for measuring cAMP levels in cells. The assay monitors cAMP production in cells 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. Perform the assay in 96-, 384- or 1536-well plates. Induce cells with a test compound to modulate cAMP levels. Lyse cells to release cAMP, and add the cAMP detection solution, which contains protein kinase A. Add Kinase-Glo Reagent to terminate the PKA reaction and detect the remaining ATP via a luciferase reaction. Read the plates using a microplate-reading luminometer. Luminescence can be correlated to the cAMP concentrations by using a cAMP standard curve.