cAMP-Glo™ Max cyclic AMP Assay
Supplier: EBRO Electronic GmbHTotal Ratings: 0
Avg. Ratings: 0.0 out of 5
The cAMP-Glo Max Assay is a homogeneous, bioluminescent and high-throughput assay to measure cyclic AMP (cAMP) levels in cells. Compounds that modulate GPCRs coupled with adenylate cyclase typically alter intracellular cAMP levels.
- Faster cAMP Detection for High-Throughput Assays
- Complete in ~30 minutes
- Scalable to 1536-well plate formats and beyond
- No interference by fluorescent compounds
The cAMP-Glo Max Assay is a homogeneous, bioluminescent and 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 cAMP-Glo Max Assay monitors cAMP levels in cells in response to the effect of agonists, antagonists or test compounds on G protein-coupled receptors (GPCRs). 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. This improved version combines the lysis and cAMP reaction buffers into the cAMP-Glo ONE Buffer. This new format streamlines the protocol and reduces the time needed to complete the assay. The new ONE Buffer is supplied at a 5X concentration, which provides increased flexibility for starting cell culture volumes. The cAMP-Glo Max Assay can be performed in 96-, 384- or 1536-well plates. The 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 (Figure 1). 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 the plates and eliminating the need for luminometers with reagent injectors.
The Max Assay can be performed in 96, 384, or 1536-well plates. The 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 with a microplate-reading luminometer. The half life for the luminescent signal is greater than four hours, providing ample time to read the plates and eliminating the need for luminometers with reagent injectors.
The Max Assay monitors cAMP levels in cells in response to the effect of agonists, antagonists, or test compounds on G protein-coupled receptors (GPCRs). 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. This improved version combines the lysis and cAMP reaction buffers into the ONE Buffer. This new format streamlines the protocol and reduces the time needed to complete the assay. The new ONE Buffer is supplied at a 5X concentration, which provides increased flexibility for starting cell culture volumes.
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