Live cell Gi - and Gs-coupled GPCR second messenger signaling

Detection of Gi and Gs -coupled GPCR second messenger signal activity has been traditionally accomplished using assays such as radioactive binding or endpoint cAMP assays that require cell lysis. Such assays measure activity at a single time point in the cellular response and do not provide kinetic information. Another option utilizes forced-coupling of Gi and Gs -coupled GPCRs to Gα16 followed by fluorescence detection of calcium flux upon agonist receptor activation. Again, this assay is sub-optimal as it does not signal through the biorelevant cAMP pathway.

Here we demonstrate endogenous receptor activity in CHO-K1 and HEK-293 cell lines stably expressing the GloSensor™ plasmid using the FLIPR System.

Read Application Note:

• Live cell Gi- and Gs -coupled GPCR second messenger signaling on the FLIPR Tetra System