Measuring dynamic and real-time cAMP levels using cADDis2, a live-cell indicator for Gs and Gi signaling
Poster Jan 25, 2017
Paul Tewson, Scott Martinka, Catherine Berlot, Thom Hughes, Anne Marie Quinn & Shane Tillo
Here, we introduce a new fluorescent cAMP indicator, cADDis2, capable of detecting dynamic changes in cAMP concentration in living cells. We have applied cADDis2 to use in a novel Gi assay that, for the first time, directly reports Gi mediated reductions in cAMP concentration, across a variety of receptors, on standard automated fluorescence plate readers.
Multiplexing cell-based assays is possible using 3D culture models that are larger and more complex than monolayers
Real-time detection methods to measure live or dead cells provide much flexibility for multiplexing
All multiplexed assay combinations should be verified using appropriate controls for each 3D cell culture model.