cAMP-Glo Assay, Promega

Supplier: Promega Corporation

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Supplier: Promega Corporation

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About this item

The cAMP-Glo Assay is a homogeneous, bioluminescent and high-throughput assay for measuring cAMP levels in cells

Simple, Bioluminescent cAMP Detection
Complete in ~45 minutes
Scalable to 1536-well plate formats and beyond
No interference by fluorescent compounds

The cAMP-Glo Assay is a homogeneous, bioluminescent and high-throughput assay for measuring cAMP levels in cells. The cAMP-Glo 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. The cAMP-Glo 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 to release cAMP, then the cAMP detection solution, which contains protein kinase A, is added. 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. Luminescence can be correlated to the cAMP concentrations by 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.

Plates are read using a microplate reading luminometer. The assay offers excellent signal-to-noise ratios. Store at –20°C (–4°F).

The cAMP-Glo assay can be performed in 96, 384, or 1536-well plates and has an approximate completion time of 45 minutes. The procedure involves inducing cells with a test compound to modulate cAMP levels. After induction, cells are lysed to release cAMP. The cAMP detection solution, which contains protein kinase A, is then added. Lastly, the Kinase-Glo™ reagent is added to terminate the PKA reaction and detect the remaining ATP via a luciferase reaction.

Ordering information: Assay sizes are determined by using approximately 60 µL final volume per well in 384-well plates. Assay includes 100 µL of cAMP (1 mM), 5 mL of cAMP-GLO Lysis buffer, 8 mL of cAMP-GLO Reaction buffer, 16 µL of Protein Kinase A, one bottle of Kinase-GLO Substrate (lyophilized), and 10 mL of Kinase-GLO Buffer.

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