2012 Promega catalogue
2012 Promega catalogue
2012 Promega catalogue
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Life<br />
Science<br />
Catalog<br />
<strong>2012</strong><br />
Worldwide Contact List<br />
Section<br />
Contents<br />
Table of<br />
Contents<br />
Cell Signaling<br />
GPCR Assays<br />
48<br />
cAMP-Glo Assay<br />
Product Size Cat.# Price ($)<br />
cAMP-Glo Assay 300 assays V1501 517.00<br />
For Research Use Only. Not for Use in Diagnostic Procedures.<br />
3,000 assays V1502 Pls. Enq.<br />
30,000 assays V1503 Pls. Enq.<br />
Description: The cAMP-Glo Assay is a homogeneous, bioluminescent and<br />
high-throughput assay for measuring cAMP levels in cells. The cAMP-Glo<br />
Assay monitors cAMP production in cells in response to the effects of test<br />
compounds on G protein-coupled receptors (GPCR). GPCRs that couple with<br />
adenylate cyclase will increase or decrease intracellular cAMP. The assay is<br />
based on the principle that cyclic AMP (cAMP) stimulates protein kinase A<br />
(PKA) holoenzyme activity, decreasing available ATP and leading to decreased<br />
light production in a coupled luciferase reaction.<br />
The cAMP-Glo Assay can be performed in 96-, 384- or 1536-well plates.<br />
The cells are induced with a test compound for an appropriate period of time<br />
to modulate cAMP levels. After induction, cells are lysed to release cAMP, then<br />
the cAMP detection solution, which contains protein kinase A, is added. The<br />
Kinase-Glo ® Reagent is then added to terminate the protein kinase A reaction<br />
and detect the remaining ATP via a luciferase reaction. Plates are read using a<br />
microplate-reading luminometer. Luminescence can be correlated to the cAMP<br />
concentrations by using a cAMP standard curve. The half-life for the luminescent<br />
signal is greater than 4 hours. This extended signal half-life eliminates<br />
the need for luminometers with injectors and allows batch-mode processing of<br />
multiple plates.<br />
Features:<br />
Fast and Easy to Use:<br />
• Assay can be completed in approximately 45 minutes.<br />
• Homogeneous.<br />
• Two steps following lysis of cells.<br />
Excellent Signal-to-Noise Ratios:<br />
• Best signal:background ratio of all the cAMP assays.<br />
• Signal:Background >200 (with cAMP), >15 (on cells).<br />
• Easily scalable to 1536-well plate formats and beyond.<br />
Proven Luminescent Technology:<br />
• Powered by Ultra-Glo Recombinant Luciferase.<br />
• No interference by fluorescent compounds.<br />
• Nonradioactive.<br />
Storage Conditions: Store the system at –20°C. Once prepared, the cAMP<br />
detection solution (cAMP-Glo Reaction Buffer with Protein Kinase A) should<br />
not be frozen. Once prepared, the Kinase-Glo ® Reagent should be dispensed<br />
into aliquots and stored at –20°C. See the product label for the expiration<br />
date.<br />
Protocol Part#<br />
cAMP-Glo Assay Technical Bulletin TB357<br />
β<br />
Phosphorylated<br />
protein kinase A<br />
substrate<br />
ADP<br />
luciferin<br />
+ O 2<br />
luciferase<br />
ATP<br />
oxyluciferin<br />
+ AMP<br />
+<br />
Light<br />
G protein-coupled<br />
receptor<br />
γ<br />
P<br />
cAMP<br />
Protein kinase A<br />
substrate<br />
For complete and up-to-date product information visit: www.promega.com/catalog<br />
β<br />
γ<br />
C C<br />
α<br />
G protein<br />
Ligand<br />
R R<br />
Active protein<br />
kinase A<br />
GDP<br />
Inactive<br />
adenylate<br />
cyclase<br />
cAMP<br />
cAMP<br />
Active<br />
adenylate<br />
cyclase<br />
α<br />
R R<br />
C C<br />
GTP<br />
ATP<br />
Inactive protein<br />
kinase A holoenzyme<br />
Schematic diagram of cAMP production in cells and the cAMP-Glo<br />
Assay.<br />
6127 MA
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