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Monitor GPCR Modulation of Cellular cAMP with an HTS Bioluminescence-Based Assay

Said A. Goueli, Kevin Hsaio, and Jolanta Vidugiriene
Promega Corporation
Publication Date: 2007

Abstract

The cAMP-Glo™ Assay is a bioluminescence assay for monitoring changes in intracellular cAMP concentrations with high sensitivity (30 fmol ± 5 SEM cAMP/well) and reproducibility (Z´ >0.8). This assay can be used to measure cAMP concentration in a single-tube or high-throughput-screening (HTS) format and is less prone to compound interference than fluorescence-based assays. We have used the cAMP-Glo™ Assay to generate EC50 values for agonists and IC50 values for antagonists that modulate GPCRs that alter adenylate cyclase activity (Gas, and Gai).The cAMP-Glo™ Assay is compatible with adherent, suspension, and frozen cells, offering flexibility and convenience to the researcher, and the cAMP-Glo™ Assay can be used to determine cAMP concentration in tissue extracts.

Promega Notes 97, 24–27.

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