GloSensor cAMP Assay An Intracellular Biosensor for cAMP Using Firefly Luciferase...
GloSensor cAMP Assay An Intracellular Biosensor for cAMP Using Firefly Luciferase Scientific Poster
Brock Binkowski, Frank Fan, Pete Stecha, Braeden Butler and Keith Wood
Promega Corporation, 2800 Woods Hollow Rd., Madison, WI 53711
Cell-based assays provide biologically relevant context for drug screening and have increased in importance in recent years. Bioluminescence is uniquely suited for developing cell-based assays, especially for high-throughput screening, owing to the inherent high sensitivity, wide dynamic range and low susceptibility to compound interference. Here we present the use of a novel biosensor construct composed of a genetic fusion of a cAMP binding domain to a circularly permuted form of firefly luciferase. This biosensor can be used to rapidly interrogate changes in the concentration of intracellular cAMP using a live cell, nonlytic assay format. This format enables direct screens for allosteric modulators of Gs- and Gi-coupled 7-TM receptors and improved hit identification through multiple measurements. Moreover, the enhanced sensitivity of the assay allows detection of inverse agonists and the activation of Gi-coupled receptors in the absence of forskolin.
- Part# PS082
- Printed in USA.