Promega Corporation

Advances In Bioluminescent Assays for G-Protein-Coupled Receptors Scientific Poster

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Abstract

Frank Fan, Aileen Paguio, Braeden Butler, Brock Binkowski, Pete Stecha and Keith Wood
Promega Corporation, 2800 Woods Hollow Rd Madison, WI 53711

Bioluminescent assays are uniquely suited for high throughput screening due to their inherent high sensitivity, wide dynamic range and low susceptibility to compound interference. G-protein-coupled receptors constitute a major target class for the development of new drugs. The activity of these receptors can be monitored via readouts at various stages of the signal transduction cascade. Traditional bioluminescent GPCR assay involves using luciferase as a genetic reporter linked to appropriate response elements. Recent improvements in reporter technology have enhanced the dynamics, sensitivity, and reliability of these assays. A new assay allows quantitation of cAMP in cell lysates through its regulation of protein kinase A activity, where the ATPase activity can be detected using bioluminescence. Another novel assay format based on a genetically encoded luciferase biosensor allows direct detection of cAMP within living cells. Comparison studies of these assays using HEK293 cells expressing dopamine receptor D1 are presented. Together, these bioluminescent assays provide more tools for GPCR drug discovery.

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  • Part# PS054
  • Printed in USA.

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