Measuring dynamic and real-time cAMP levels using cADDis

Real-Time cAMP Detection

Here, we introduce a new fluorescent cAMP sensor, cADDis, capable of detecting dynamic changes in cAMP concentration in living cells. We have applied cADDis 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.

cADDis2 cAMP Indicator

• Genetically encoded, single fluorescence emission
• Packaged in a variety of viral vectors for use in any cell type
• Decrease in fluorescence signal upon binding cAMP
• Easy to use on standard fluorescence microscopes and automated fluorescence plate readers

Green fluorescent cADDis sensor in HEK293 cells responding to isoproterenol, a selective agonist for the endogenous β2 adrenergic receptor.

A New Way to Detect Gi Signaling

• Simple, No cell lysis, No FRET, Fully reversible
• Real-time detection of dynamic cAMP levels
• All data obtained on Biotek Synergy MX plate reader

First ever direct readout of Gi mediated reductions in cAMP

• Tuned expression of constitutively active Gαs raises basal levels of cAMP, without saturating cADDis
• No pre-treatment with Gs agonists, Forskolin, or IBMX
• Reveal the activity of Phosphodiesterases