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 cAMP 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 cAMP assay that detects changes in cAMP in real time. 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.
- Screen Gs or Gi-coupled GPCRs in living cells
- 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
- Target to microdomains or specific cells in mixed cultures
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
- K.M. Semesta, et al. The psychosis risk factor RBM12 encodes a novel repressor of GPCR/cAMP signal transduction. Journal of Biological Chemistry. August 2023.
- Y.J. Peng, et al. Hypoxia sensing requires H2S-dependent persulfidation of olfactory receptor 78. Science Advances. July 2023.
- S. Zhang, et al. Competition between stochastic neuropeptide signals calibrates the rate of satiation. bioRxiv. July 2023.
- E. Kitayama, et al. Functional Expression of IP, 5-HT4, D1, A2A, and VIP Receptors in Human Odontoblast Cell Line. Biomolecules. May 2023.
- D. Santana Nunez, et al. Piezo1 induces endothelial responses to shear stress via soluble adenylyl Cyclase-IP3R2 circuit. iScience. May 2023.
- S. Bitsi, et al. Divergent acute versus prolonged pharmacological GLP-1R responses in adult β cell–specific β-arrestin 2 knockout mice. Science Advances. May 2023.
- V. Bhatia, et al. Characterization of Adenylyl Cyclase Isoform 6 Residues Interacting with Forskolin. Biology. April 2023.
- H. Carr, et al. The Wnt pathway protein Dvl1 targets Somatostatin receptor 2 for lysosome-dependent degradation. Journal of Biological Chemistry. March 2023.
- I. Cattani-Cavalieri, et al. Quantitative phosphoproteomic analysis reveals unique cAMP signaling pools emanating from AC2 and AC6 in human airway smooth muscle cells. Frontiers in Physiology. February 2023.
- E. Porpiglia, et al. Elevated CD47 is a hallmark of dysfunctional aged muscle stem cells that can be targeted to augment regeneration. Cell Stem Cell. December 2022. (bioRxiv)
- J. Janetzko, et al. Membrane phosphoinositides regulate GPCR-β-arrestin complex assembly and dynamics. Cell. November 2022.
- N. Saito, et al. Gαs-Coupled CGRP Receptor Signaling Axis from the Trigeminal Ganglion Neuron to Odontoblast Negatively Regulates Dentin Mineralization. biomolecules. November 2022.
- J. Xu, et al. An evolutionarily conserved olfactory receptor is required for sex differences in blood pressure. bioRxiv. November 2022.
- B. Barsi-Rhyne, et al. Discrete GPCR-triggered endocytic modes enable β-arrestins to flexibly regulate cell signaling. eLife. October 2022. (bioRxiv)
- S. A. Dai, et al. State-selective modulation of heterotrimeric Gαs signaling with macrocyclic peptides. Cell. September 2022.
- J. H. Cho, et al. Islet primary cilia motility controls insulin secretion. Science Advances. September 2022. (bioRxiv)
- E. Blythe, M. von Zastrow. A discrete mode of endosomal GPCR signaling that does not require β-arrestins. bioRxiv. September 2022.
- ER McGlone, et al. Hepatocyte cholesterol content modulates glucagon receptor signaling. Molecular Metabolism. September 2022.
Posters: cADDis cAMP Assay
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
GPCR Biology
Increase your understanding of drug effects and GPCR biology with bright fluorescent assays for Gs, Gi, and Gq signaling in living cells.
cADDis cAMP Assays
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#D0200G Green Down cADDis cAMP Assay Kit
$695.00 – $2,395.00 -
#U0200G Green Up cADDis cAMP Assay Kit
$695.00 – $2,395.00 -
#X0200G Green Gi cADDis cAMP Assay Kit
$695.00 – $2,395.00 -
#U0200R Red Up cADDis cAMP Assay Kit
$695.00 – $2,395.00 -
#D0201G Green Cilia-targeted cADDis cAMP Assay Kit
$695.00 – $1,495.00 -
#D0211G Ratiometric Cilia-Targeted cADDis cAMP Assay Kit
$695.00 – $1,495.00 -
#D0231G Green Membrane-Targeted FMP cADDis cAMP
$695.00 – $1,495.00 -
#D0241G Green Membrane-Targeted fS15 cADDis cAMP
$695.00 – $1,495.00 -
#U0241R Red Membrane-Targeted fS15 cADDis cAMP
$695.00 – $1,495.00 -
#U0205G Green Upward cADDis cAMP (Promoter CAG)
$695.00 – $2,395.00
