Description
Bioluminescent Detection of cAMP to Monitor GPCR Activity
The cAMP-Glo™ Max Assay is a homogeneous, bioluminescent, high-throughput assay to measure cyclic AMP (cAMP) levels in cells. Compounds that modulate GPCRs coupled with adenylate cyclase typically alter intracellular cAMP levels. The assay monitors cAMP levels in cells in response to the effect of agonists, antagonists or test compounds on G protein-coupled receptors (GPCRs), and 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.
Improved Format for Better Performance
This improved version of the cAMP-Glo™ Assay combines the lysis and cAMP reaction buffers into the cAMP-Glo™ ONE Buffer, streamlining the protocol and speeding up the assay. The ONE Buffer is supplied at 5X concentration, which provides increased flexibility for starting cell culture volumes.
The cAMP-Glo™ Max Assay allows easy quantitation of cAMP levels in broad range of cell types, and can be performed in 96-, 384- or 1536-well plates. Cells are induced with a test compound for an appropriate period of time to modulate cAMP levels. After induction, cells are lysed and the cAMP released stimulates protein kinase A in the reagent. The Kinase-Glo® Reagent is then added to terminate the PKA reaction and detect the remaining ATP via a luciferase reaction. Plates are read using a microplate-reading luminometer. The half-life for the luminescent signal is greater than 4 hours, providing ample time to read plates and eliminating the need for luminometers with reagent injectors.
Cat.# V1681 contains sufficient reagents for 2 plates (96- or 384-well format). Cat.# V1682 contains sufficient reagents for 20 plates (96- or 384-well format). Cat.# V1683 contains sufficient reagents for 200 plates (96- or 384-well format).
cAMP production in cells and the cAMP-Glo™ Max Assay.
Schematic diagram of cAMP production in cells and the cAMP-Glo™ Max Assay.
