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
Example publications using these tools:
- ER McGlone, et al. Hepatocyte cholesterol content modulates glucagon receptor signaling. Molecular Metabolism. September 2022.
- N. Senese, et al. Antidepressants produce persistent Gαs associated signaling changes in lipid rafts following drug withdrawal. Molecular Pharmacology. May 2021.
Nucleus-targeted cAMP Assays
Cilia-targeted cAMP Assays
Publications using these tools:
- J. Hansen, et al. A cAMP signalosome in primary cilia drives gene expression and kidney cyst formation. EMBO Reports. June 2022.
- C. Wu, et al. Discovery of ciliary G protein-coupled receptors regulating pancreatic islet insulin and glucagon secretion. Genes & Development. August 2021.
- R. Sherpa et al. Sensory primary cilium is a responsive cAMP microdomain in renal epithelia. Scientific Reports. April 2019
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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!