Detect Gs or Gi signaling with cADDis, our cAMP Assay
- Use cADDis to measure cAMP through Gs or Gi signaling in real time with a single sensor
- Plate reader or imaging system compatible
Downward cADDis starts out bright and decreases in fluorescence when cAMP levels increase. This cAMP assay is easily detected on fluorescence plate readers and helps with imaging cells that have low expression efficiency.
Upward cADDis starts out dim and increases in fluorescence when when cAMP levels increase. This cAMP assay is easily detected on fluorescence plate readers or imaging systems.
cADDis cAMP Assay Performance
Off the shelf kits include the cAMP sensor in BacMam, a BSL-1 viral vector for efficient delivery to most cell types. Purified BacMam, AAV, Lenti vectors by request.
- Easily detectable on plate readers or imaging systems
- Controllable expression in BacMam vector
- High signal-to-noise ratio
Red DAG sensor multiplexed with a green cAMP sensor indicates dose-dependent signaling via a calcitonin receptor.
Multiplex Assays
- Combine red and green sensors to measure multiple GPCR pathways simultaneously
A Robust and Versatile cAMP Assay Kit:
cADDis is a genetically-encoded fluorescent cAMP assay that detects changes in cAMP in real time.
- Screen Gs or Gi-coupled GPCRs in living cells
- Robust expression in practically any cell type
- Target to microdomains or specific cells in mixed cultures
Simple protocol:
Provided online is a simple cAMP assay protocol, with no need for co-factors, cell lysis, or complex liquid handling.
- Text and video protocols online
- No lysis or complex liquid handling
- Single channel, non-FRET signal
Check out this Live Cell Imaging video from BioTek Instruments: Kinetic Characterization of Gs- and Gi-dependent regulation of cAMP.
It’s a brief animated presentation demonstrating expression & kinetic monitoring of our fluorescent cADDis cAMP Assay in live cells.
Recent Publications
- J. Yu et al. N-3 polyunsaturated fatty acids promote astrocyte differentiation and neurotrophin production independent of cAMP in patient-derived neural stem cells. Mol. Psychiatry. June 2020.
- Nunez, F.J., Johnstone, T.B. et al. Glucocorticoids rapidly activate cAMP production via Gαs to initiate non‐genomic signaling that contributes to one‐third of their canonical genomic effects FASEB Journal, Dec. 2019
- Nunez, F.J., Schulte, N.A., Fogel, D.M., et al. Agonist-specific desensitization of PGE2-stimulated cAMP signaling due to upregulated phosphodiesterase expression in human lung fibroblasts Naunyn-Schmiedeberg’s Arch Pharmacol Dec. 2019
- P. Salas, et al. Glucocorticoids and Myosin5b loss-of-function induce heightened PKA signaling in addition to membrane traffic defects MBoC Oct, 2019
- R. Lee, et al. Bitter taste receptors stimulate phagocytosis in human macrophages through calcium, nitric oxide, and cyclic-GMP signaling bioRxiv Oct, 2019
- R. Lee, et al. Inverse regulation of secretion and inflammation in human airway gland serous cells by neuropeptides upregulated in allergy and asthma bioRxiv May 10, 2019
- T. Baldwin, et al. Insights into the Regulatory Properties of Human Adenylyl Cyclase Type 9 Molecular Pharmacology. April, 2019
- T. Togo Autocrine purinergic signaling stimulated by cell membrane disruption is involved in both cell membrane repair and adaptive response in MDCK cells Biochem & Biophysical Research Comm. March, 2019
- R. Ostrom, et al. Transforming Growth Factor-β1 Decreases β2-Agonist–induced Relaxation in Human Airway Smooth Muscle ATSJournas, Dec 2018
- X. Chen, et al. Phenylephrine, a common cold remedy active ingredient, suppresses uterine contractions through cAMP signalling. Scientific Reports Aug. 2018.
- T. Buranda, et al. A High-Throughput Flow Cytometry Screen Identifies Molecules that Inhibit Hantavirus Cell Entry. SLAS Discovery. April 2, 2018.
- N. H. Wray, et al. NMDAR-independent, cAMP-dependent antidepressant actions of ketamine. Molecular Psychiatry. April 2, 2018.
- H. Zou, et al. PDE8: A Novel Target in Airway Smooth Muscle. American Journal of Respiratory Cell and Molecular Biology. Vol. 58, No. 4. April 01 2018.
- T.B. Johnstone, et al. PDE8 is Expressed in Human Airway Smooth Muscle and Selectively Regulates cAMP Signaling by β2AR-AC6. American Journal of Respiratory Cell and Molecular Biology. Dec. 2017.
- K.A. McCrink, et al. β-Arrestin2 Improves Post-Myocardial Infarction Heart Failure via Sarco(endo)plasmic Reticulum Ca2+-ATPase-Dependent Positive Inotropy in Cardiomyocytes. Hypertension. Nov. 2017.
- J. Almaça, et al. Human beta cells produce and release serotonin to inhibit glucagon secretion from alpha cells.Cell Reports, Volume 17, Issue 12, 2016.
- P. Tewson, et al. New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories. J Biomol Screen Dec. 11, 2015. Request Full Text PDF
- P. Tewson, et al. Assay for Detecting Gαi-Mediated Decreases in cAMP in Living Cells. SLAS. July 10, 2018. Request Full Text PDF
- Y.A.B. Sierra, et al. Potassium channel-based optogenetic silencing. Nature Communications Nov. 5 2018.
- M. Thomas, et al. Toward a High Throughput Screen for Voltage Sensors. 2018.
- K. Hilgendorf, et al. Omega-3 Fatty Acids Activate Ciliary FFAR4 to Control Adipogenesis Cell, November, 2019
- R. Sherpa, et al. Sensory primary cilium is a responsive cAMP microdomain in renal epithelia Scientific Reports, April 25, 2019
- B.S. Moore, et al. Cilia have high cAMP levels that are inhibited by Sonic Hedgehog-regulated calcium dynamics. PNAS Nov. 15, 2016.
cADDis – cAMP Assay Posters
Measuring dynamic and real-time cAMP levels using cADDis, a live-cell indicator for Gs and Gi signaling
PDE Specificity Illuminated: Monitoring cGMP and cAMP Levels in Living Cells
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