Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

The Equilibrium Binding Constant and Binding Strength02:18

The Equilibrium Binding Constant and Binding Strength

The equilibrium binding constant (Kb) quantifies the strength of a protein-ligand interaction. Kb can be calculated as follows when the reaction is at equilibrium:
Ligand Binding Sites02:40

Ligand Binding Sites

Proteins are dynamic macromolecules that carry out a wide variety of essential processes; however, the activities of most proteins depend on their interactions with other molecules or ions, known as ligands.
Protein-ligand interactions are quite specific; even though numerous potential ligands surround a cellular protein at any given time, only a particular ligand can bind to that protein. Moreover, a ligand binds only to a dedicated area on the surface of the protein, known as the...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Platelets and allergic inflammation.

Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology·2014
Same author

Induction of platelet aggregation after a direct physical interaction with diesel exhaust particles.

Journal of thrombosis and haemostasis : JTH·2012
Same author

Cytokines in airway inflammation.

The international journal of biochemistry & cell biology·2000
Same author

Murine models of inflammation: role of CD23.

Allergy·2000
Same author

Squalamine inhibits angiogenesis and solid tumor growth in vivo and perturbs embryonic vasculature.

Cancer research·1998
Same author

Characterization of functional activity of chemokine receptors using the Cytosensor Microphysiometer.

Methods in enzymology·1997
Same journal

Simultaneous Ca<sup>2+</sup> Imaging and Optogenetic Stimulation of Cortical Astrocytes in Adult Murine Brain Slices.

Current protocols in neuroscience·2020
Same journal

Automated Two-Chamber Operon ID/ED Task for Mice.

Current protocols in neuroscience·2020
Same journal

A Guide to Fluorescence Lifetime Microscopy and Förster's Resonance Energy Transfer in Neuroscience.

Current protocols in neuroscience·2020
Same journal

Development, Screening, and Validation of Camelid-Derived Nanobodies for Neuroscience Research.

Current protocols in neuroscience·2020
Same journal

Recombinant Antibodies in Basic Neuroscience Research.

Current protocols in neuroscience·2020
Same journal

Automated Quantification of Mitochondrial Fragmentation in an In Vitro Parkinson's Disease Model.

Current protocols in neuroscience·2020
See all related articles

Related Experiment Video

Updated: Jul 5, 2026

Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects
13:57

Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects

Published on: February 18, 2014

Ligand characterization using microphysiometry.

S Pitchford1

  • 1Molecular Devices Corporation, Sunnyvale, California, USA.

Current Protocols in Neuroscience
|April 23, 2008
PubMed
Summary
This summary is machine-generated.

This study details using a Cytosensor microphysiometer to characterize G protein-coupled receptor activity. It optimizes ligand exposure and recovery times for accurate muscarinic M1 receptor agonist and antagonist analysis.

More Related Videos

PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions
10:58

PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions

Published on: July 27, 2017

Related Experiment Videos

Last Updated: Jul 5, 2026

Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects
13:57

Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects

Published on: February 18, 2014

PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions
10:58

PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions

Published on: July 27, 2017

Area of Science:

  • Pharmacology
  • Cellular Biology
  • Biophysics

Background:

  • G protein-coupled receptors (GPCRs) are crucial drug targets.
  • Muscarinic M1 receptors are implicated in various physiological processes.
  • Accurate functional characterization of receptor ligands is essential for drug discovery.

Purpose of the Study:

  • To describe a method for functional characterization of muscarinic M1 receptor ligands using a Cytosensor microphysiometer.
  • To optimize experimental parameters for accurate EC50 and pA2 value determination.
  • To demonstrate the versatility of the Cytosensor for both adherent and suspension cell cultures.

Main Methods:

  • Utilizing a Cytosensor microphysiometer to measure cellular responses to ligand stimulation.
  • Generating concentration-response profiles for agonists and antagonists.
  • Optimizing ligand exposure duration and inter-ligand recovery time to mitigate receptor desensitization.
  • Calculating EC50 for agonists and pA2 values for antagonists.

Main Results:

  • Established a protocol for functional characterization of muscarinic M1 receptor ligands.
  • Determined optimal conditions for ligand exposure and recovery to ensure reliable measurements.
  • Demonstrated the ability to quantify agonist potency (EC50) and antagonist affinity (pA2).
  • Validated the use of the Cytosensor microphysiometer with both adherent (M1WT3) and suspension cell cultures.

Conclusions:

  • The Cytosensor microphysiometer provides a robust platform for functional characterization of GPCRs, specifically the muscarinic M1 receptor.
  • Optimized protocols enhance the accuracy and reliability of ligand potency and affinity measurements.
  • This method is adaptable for various cell types, facilitating broader applications in pharmacological research.