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GPCR Assay Services
Compound Profiling
- Measure Gs, Gi, Gq, and β-arrestin activity in real time.
- Quantify the dynamics of signaling, including signaling duration, initial rate, and desensitization kinetics.
- Dose-response curves from initial rate, peak activation, and signal plateau.
- Quantify partial or biased agonism.
- Estimate efficacy, potency and desensitization rate.
Case study: Profiling µ-opioid receptor agonists in living cells
with biosensors for cAMP and β-arrestin.
![Fig-2_camp](https://montanamolecular.com/wp-content/uploads/2023/02/Fig-2_camp-1.jpg)
Time course data from cAMP cADDis assay fit to kinetic equations. Analysis estimates initial rate of signaling (dashed line) at each dose of morphine at the µ-opioid receptor.
![cAMP-fig 1](https://montanamolecular.com/wp-content/uploads/2023/02/cAMP-fig-1.jpg)
cADDis biosensor reveals distinct temporal profiles of cAMP signaling.
![cAMP-fig-3](https://montanamolecular.com/wp-content/uploads/2023/03/cAMP-fig-3.jpg)
CRCs for 5 µ-opioid receptor agonists using the initial rate of activation.
![MOR---kinetic](https://montanamolecular.com/wp-content/uploads/2023/02/MOR-kinetic-1.jpg)
![arrestin-fig-1](https://montanamolecular.com/wp-content/uploads/2023/02/arrestin-fig-1-3.jpg)
![arrestin-fig-2](https://montanamolecular.com/wp-content/uploads/2023/02/arrestin-fig-2.jpg)
Summary of results at the µ-opioid receptor
- All compounds generate cAMP at a similar rate, except buprenorphine (a partial agonist).
- Small molecules recruit arrestin more slowly than peptides.
- The ratio of initial rates can be used to quantify bias for cAMP over arrestin – small molecules are biased for cAMP.
![arrestin-table](https://montanamolecular.com/wp-content/uploads/2023/02/arrestin-table-1.jpg)
Video: Description of our Assay Services
Data Analysis
Time course equations for a variety of curve shapes entered into GraphPad Prism as a plug-in.
Parameters include:
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- Initial rate of signal generation
- Fade (desensitization) rate
- Steady-State signal level
- Peak level and peak time
References
![data-analysis-fig-1](https://montanamolecular.com/wp-content/uploads/2023/02/data-analysis-fig-1.jpg)
Rise and fall to steady-state curve
![data-analysis-fig-2](https://montanamolecular.com/wp-content/uploads/2023/02/data-analysis-fig-2.jpg)
Rise to steady-state curve
Quantifying GPCR Signaling and Desensitization Kinetics
![arrestin-recycling](https://montanamolecular.com/wp-content/uploads/2023/02/arrestin-recycling-1.jpg)
Waveform data fitted to equations to quantify:
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- Signal generation rate
- Desensitization rate
- Signal clearance rate
- Differences between GPCRs in signaling mechanisms and rates
![Oxytocin-&-M1-DAG](https://montanamolecular.com/wp-content/uploads/2023/02/Oxytocin-M1-DAG.jpg)
Assay Optimization Services
Our GPCR Discovery Team can kick start your in-house discovery efforts with our assay optimization services. You will receive your receptor in a BacMam delivery vector to ensure consistent, controllable receptor expression, an optimized protocol, and biosensor assay starter kit. Choose from GPCR assays for β-arrestin, cAMP, DAG, PIP2, and Calcium, or ask us about our biosensors for cGMP, Cell Stress, and SARS-CoV-2 protease inhibitors.
Get A Quote or Inquire about a Statement of Work for your Project
We don’t just run assays,
we have developed a full suite of the best in class, fluorescent biosensor assay kits for
do-it-yourselfers.