Abstract: The poster also demonstrates endogenous receptor activity in CHO-K1 and HEK-293 cell lines stably expressing the GloSensor plasmid. In addition, transfected Gs- and Gi-coupled receptor activity are measured from cell lines with stably transfected GPCR receptors and transient transfection of the GloSensor plasmid. Multiplexing Gq- and Gs-coupled GPCR assays was shown to be possible using the GloSensor cAMP assay and FLIPR® Calcium 5 Kit in the same well. Combined with the GloSensor cAMP Assay, the FLIPR® Tetra system delivers the complete solution for kinetic screening of the major classes of GPCR subtypes.Summary: Detection of Gs- and Gi-coupled GPCR second messenger signal activity has been traditionally accomplished using endpoint assays such as radioactive binding or cAMP assays that require cell lysis. This poster demonstrates the use of the modified luminescent firefly luciferase-based Promega GloSensor™ cAMP Assay on the FLIPR® Tetra system to enable detection of cAMP mediated Gs- and Gi-coupled GPCR activity in a true kinetic assay.References:
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