cADDis: Live Cell Assays for cAMP
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
Measure cAMP through Gs or Gi Signaling in Real Time with a Single Sensor
- Quantify kinetics of cyclic AMP signaling in living cells.
- Measure cAMP for many hours with minimal loss of sensor signal.
- Robust expression in practically any cell type.
Quantify cAMP Kinetics Precisely
- Changes over time reflect biological signaling mechanisms (rather than technical artifacts like sensor decay).
- Quantify signaling mechanisms by curve fitting time course data (see here for example report).
- In the example below we quantify the signal generation rate, signal decline resulting from desensitization, and late stage decline (likely resulting from peptide ligand degradation).
Your endpoint assays are missing important information. Measuring kinetics with our live-cell assays can reveal it.
- Signaling dynamics of multiple drug compounds
- Different dynamics of cAMP generation by β2 adrenergic receptor agonists
- Clenbuterol and salbutamol generate cAMP more slowly and clenbuterol desensitizes receptor more slowly
A Robust & Versatile cAMP Assay Kit
- cADDis is a genetically-encoded fluorescent cyclic AMP assay that detects changes in cAMP in real time. Off the shelf kits include the cADDis sensor in BacMam, a BSL-1 viral vector for efficient delivery to most cell types. Purified BacMam, AAV, Lenti vectors by request.
- Screen Gs or Gi-coupled GPCRs in living cells
- Target to microdomains or specific cells in mixed cultures
- Robust expression in practically any cell type
- Easily detectable on plate readers or imaging systems
- Controllable expression in BacMam viral vector
- High signal-to-noise ratio
Simple Protocol
- Minimal liquid handling
- No cell lysis
- 96 or 384 well compatible
- Single channel, non-FRET signal
GPCR Assay Services
Go from hit to lead faster with compound profiling assays that capture GPCR signaling dynamics in living cells. Our unique biosensor-based discovery platform quantifies multiple signaling parameters in real time to enable decisions based on clinically-relevant data. Our fast turnaround time helps reduce the time to the clinic. We will perform experiments to advance your projects and programs, for example by optimizing assays, profiling compounds, exploring mechanisms and kinetically characterizing signaling dynamics.
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
- L. Coassolo, et al. Prohormone cleavage prediction uncovers a non-incretin anti-obesity peptide. Nature. March 2025.
- R. Schuck, et al. Cholesterol inhibits assembly and oncogenic activation of the EphA2 receptor. Nature Communications Biology. March 2025.
- E. Blythe, et al. Endocytosis sculpts distinct cAMP signal transduction by endogenously coexpressed GPCRs. bioRxiv preprint. February 2025.
- S. Orfanos, et al. The differential effects of cAMP mobilizing agents on inhibition of TGF-β-induced extracellular matrix and growth factor expression in human lung fibroblasts. Authorea. February 2025.
- A. Klein, et al. Inhibition of adenylyl cyclase 1 (AC1) and exchange protein directly activated by cAMP (EPAC) restores ATP-sensitive potassium (KATP) channel activity after chronic opioid exposure. bioRxiv. February 2025.
- I. Davies, et al. Chronic GIPR agonism results in pancreatic islet GIPR functional desensitisation. Molecular Metabolism. January 2025.
- N. Patel, et al. Myosin VI drives arrestin-independent internalization and signaling of GPCRs. Nature Communications. December 2024.
- C. Bonner, et al. Metabolic State Determines Central and Peripheral Mechanisms of Liraglutide-Enhanced Insulin Secretion. Research Square. December 2024.
- M. Kimura, et al. Intracellular cAMP signaling-induced Ca2+ influx mediated by calcium homeostasis modulator 1 (CALHM1) in human odontoblasts. European Journal of Physiology. November 2024.
- A. Ali, et al. Comparative Effects of GLP-1 and GLP-2 on Beta-Cell Function, Glucose Homeostasis and Appetite Regulation. Biomolecules. November 2024.
- A. Hamilton, et al. Nicotinic signaling stimulates glucagon secretion in mouse and human pancreatic α-cells. Diabetes. October 2024.
- LE Eid, et al. In vivo functional profiling and structural characterisation of the human Glp1r A316T variant. bioRxiv. October 2024.
- L. Ripoll, et al. Spatial organization of adenylyl cyclase and its impact on dopamine signaling in neurons. Nature Communications. September 2024.
- R. Nassini, et al. Targeting the Schwann Cell EP2/cAMP Nanodomain to Block Pain but not Inflammation. bioRxiv. September 2024.
- S. Maghsoudi, et al. Adenylyl Cyclase Isoform 6 in the Pulmonary Artery Is Inhibited by Hypoxia via Cysteine Nitrosylation. American Journal of Respiratory Cell and Molecular Biology. August 2024.
- Y. Manchange, et al. Engineered mini-G proteins block the internalization of cognate GPCRs and disrupt downstream intracellular signaling. Science Signaling. July 2024.
Posters: cADDis cAMP Assay
Quantifying the kinetics of endocrine/metabolic GPCR signalling using high performance biosensors and a curve fitting platform
Montana Molecular has developed biosensors of GPCR signaling with unprecedented performance, enabling sub-second read frequency, total read times of over five hours, and miniaturization to 384-well format, for Gs, Gi, Gq and arrestin pathways. Signaling dynamics are quantified using simple curve fitting methods using a free plug-in to the commonly-used program GraphPad Prism. Here these technologies are applied to quantify the signaling and desensitization dynamics of endocrine and metabolic GPCRs, to provide high quality, actionable results to advance research and drug discovery programs.
GPCR signaling kinetics in pain and OUD: A platform for measuring and analyzing the time course of signal transduction
- The timing (dynamics) of GPCR signaling impacts the physiological function and therapeutic activity of ligands activating the receptors.
- Bright fluorescent biosensors enable continuous recording of GPCR signaling in live cells, with high read frequency (seconds) for long durations (hours)
- Kinetic parameters can be extracted from the waveform data by curve fitting using a plug-in for the popular program GraphPad Prism
- This platform is applied to quantify the kinetics of partial agonism and bias at the μ-opioid receptor, and the dynamics of arrestin recruitment to multiple GPCRs.
GPCR Biology
Increase your understanding of drug effects and GPCR biology with bright fluorescent assays for Gs, Gi, and Gq signaling in living cells.
All cADDis cAMP Assays
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#D0201G Green Cilia-targeted cADDis cAMP Assay Kit
$995.00 -
#D0211G Ratiometric Cilia-Targeted cADDis cAMP Assay Kit
$995.00 -
#D0221G Green Nuclear-Targeted cADDis cAMP
$995.00 -
#U0221R Red Nucleus-Targeted cADDis cAMP
$995.00 -
#D0231G Green Membrane-Targeted FMP cADDis cAMP
$995.00 -
#D0241G Green Membrane-Targeted fS15 cADDis cAMP
$995.00 -
#U0241R Red Membrane-Targeted fS15 cADDis cAMP
$995.00 -
#U0263G AKAP Targeted cADDis cAMP Sensor – AKAP1
$895.00 -
#U0264G AKAP Targeted cADDis cAMP Sensor – AKAP79
$895.00 -
#U0266G AKAP Targeted cADDis cAMP Sensor – AKAP12
$895.00 -
#U0267G AKAP Targeted cADDis cAMP Sensor – smAKAP
$895.00 -
#U0262G AKAP Targeted cADDis cAMP Sensor – AKAP18b
$895.00 -
#U0261G AKAP targeted cADDis cAMP Sensor – AKAP18a
$895.00 -
#D0250G Green Down cADDis cAMP Assay Kit (Big Sky)
$895.00 – $2,995.00 -
#U0250G Green Up cADDis cAMP Assay Kit (Big Sky)
$895.00 – $2,995.00 -
#U0250R Red Up cADDis cAMP Assay Kit (Big Sky)
$895.00 – $2,995.00 -
#X0250G Green Gi cADDis cAMP Assay Kit (Big Sky)
$895.00 – $2,995.00
