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
Direct, Real-Time Gi Detection
![](https://montanamolecular.com/wp-content/uploads/2017/01/Quinpirole-D2-receptor.jpg)
![](https://montanamolecular.com/wp-content/uploads/2017/01/Gi-Agonist.jpg)