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Promega™ GloSensor™ cAMP Assay

Provides an extremely sensitive and easy-to-use format for the interrogation of overexpressed or endogenous G-protein coupled receptors (GPCRs) that signal via changes in the intracellular concentration of cAMP

Manufacturer: promega™  E1261

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GloSensor cAMP Reagent is required for use with this assay per the GloSensor Limited Use Label License.

Catalog No. PRE1261

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Description & Specifications


Detection Method Luminescence
Format Fluid
Includes 2 vials of 2 × 106; GloSensor cAMP HEK293 Cell Line
pH 7.5
For Use With (Application) Full and partial agonists and antagonists as well as inverse agonists and allosteric modulators
Quantity 2 Vials
Storage Requirements -140°C

The assay uses genetically encoded biosensor variants with cAMP binding domains fused to mutant forms of Photinus pyralis luciferase. Upon binding to cAMP, conformational changes occur that promote large increases in light output. Following pre-equilibration with substrate, cells transiently or stably expressing a biosensor variant can be used to assay GPCR function using a live-cell, nonlytic assay format, enabling simple kinetic measurements of cAMP accumulation or turnover in living cells.

  • Broad dynamic range with up to 1000-fold changes in light output obtained
  • Easy monitoring of cAMP in live cells enables a more complete analysis of receptor biology
  • High sensitivity assay permits easy detection of low-abundance, endogenous receptors
  • Direct detection of Gi-coupled receptor activation and inverse agonist activity in the absence of artificial stimulation by compounds such as forskolin provides increased biological relevance
  • Live-cell, nonlytic assay format greatly facilitates HTS/uHTS

Choice of Two Biosensor Plasmids

  • pGloSensor™-22F cAMP Plasmid (Mfr. No. E2301) is the first choice for most applications
  • Following cell-free expression in vitro, the biosensor encoded by pGloSensor-22F cAMP Plasmid shows an increased EC50 for activation together with increased signal-to-background ratio at cAMP saturation relative to the version encoded by the pGloSensor-20F cAMP construct
  • In general, similar relationships have been observed between the two constructs when their performance is compared in living cells
  • pGloSensor-20F cAMP Plasmid (Mfr. No. E1171) encodes a version of the biosensor that performs well in HEK293 cells at 37°C Luminescence from the pGloSensor-22F cAMP Plasmid construct can be more difficult to detect at physiologic temperatures

Screen for full and partial agonists and antagonists as well as inverse agonists; Screen for allosteric modulators; Profile Gs- and Gi-coupled receptor biology; Characterize cAMP changes in live cells; Assay pharmacology of compounds in live cells