Select Page

Targeted cADDis: Live Cell Assays for cAMP in Sub-cellular Domains

 There is a growing body of evidence that cAMP signaling can be quite localized. Our next-generation of cADDis cAMP assays are targeted to micro and nano domains of signaling.

  • Detect cAMP changes in living cells with targeted cADDis sensors, our cAMP Assays to detect localized signaling kinetics
  • Plate reader or imaging system compatible

Genetic targeting

Montana Molecular’s cADDis biosensor for cAMP is a cytosolic protein that measures the total level of cytoplasmic cAMP, providing robust, reproducible measurements of receptor activation and the waveform of the cellular response of the cell over minutes or hours (Hoare & Hughes, 2021). To measure cAMP signaling in specific signalsomes, new versions of cADDis were genetically targeted to effectively measure cAMP in particular subcellular compartments. 

AKAP-targeted cAMP Assays

There are many different AKAP Proteins that are localized to different regions of the cell. These AKAP Proteins bind the regulatory subunits of the PKA enzyme, but they often interact with adenyl cyclases, phosphodiesterases, and PKA substrates. This has led to the idea that they define “signalsomes” in the cell.

We have fused AKAP18 alpha, AKAP18 beta, AKAP1, AKAP79, AKAP 12, and smAKAP to our cADDis cAMP sensor. The videos below show these tools responding to isoproterenol in HEK293 cells.

Membrane-targeted cAMP assays

Flag-tagged MyrPalm and S15 motifs target the cADDis sensor to the plasma membrane. Only green is available for fMP-targeted cADDis (D0231G), both red (U0241R) and green (D0241G) fluorescent fS15-targeted cADDis are available.

Example publications using these tools:

Images showing membrane-targeted cyclic AMP sensor expression

Nucleus-targeted cAMP Assays

  • cADDis sensors which have been targeted to the nucleus with an NLS motif. Red (U0221R) and Green (D0221G) NLS-targeted sensors are available. Both respond robustly to intracellular cAMP changes.
Figure showing kinetic responses to red and green fluorescent nucleus targeted cAMP sensors

Cilia-targeted cAMP Assays

Our cilia-targeted cAMP assays were localized using a 5HT6 receptor. Green-only (D0201G) and ratiometric (D0211G) versions are available.

Publications using these tools:

Did you find what you were looking for? We’d love to hear about your project and discuss other tools that would help you get the subcellular signaling data you need. Please reach out to the team at info@montanamolecular.com!

 

 

GPCR Biology

Increase your understanding of drug effects and GPCR biology with bright fluorescent assays for Gs, Gi, and Gq signaling in living cells.

Shop Other Assay Kits by Category