Millennium Science GloSensor cAMP assay

Wednesday, 02 February, 2011 | Supplied by: Millennium Science Pty Ltd

G-protein coupled receptors (GPCRs) are an important protein family in pharmacological applications and drug discovery research as they are involved with many critical cellular signalling responses. Monitoring the response of GPCRs to a variety of agonist and antagonists is currently being investigated by academic researchers and drug discovery efforts. Traditionally, methods in cAMP quantification require cell lysis prior to detection of intracellular cAMP levels with a colorimetric readout. This method was improved by increasing sensitivity using methods such as TR-FRET and bead-based assays, though they were considered end-point assays and still required cell lysis prior to detection.

Recently, investigations have been undertaken and findings showed that, by using a bioluminescent sensor such as the GloSensor cAMP HEK293 L/9 -2 cell line and GloSensor technology, allowed for quantitative analysis of β-adrenergic receptor activity by monitoring intracellular cAMP levels. The GloSensor cAMP assay showed high sensitivity and low background signals from as little as 1000 cells/well. The live-cell capability of the assay allowed for the use of kinetic measurements required for the investigation of both agonist and antagonist.

The SynergyH4 multimode microplate reader with hybrid technology was used in this investigation and proved to be a suitable platform with its dedicated optical system for luminescence detection, allowing it to perform such applications as the live-cell GloSensor cAMP Assay. The combination of the assay and instrumentation provide a suitable solution for high-throughput detection of GPCR signal transduction.

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