Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

The Equilibrium Binding Constant and Binding Strength

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:

You might also read

Related Articles

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

Sort by

Same author

CRISPR/Cas12a2-Mediated Ultrasensitive Assay for Rapid Detection of H1N1 Influenza Virus RNA.

ACS omega·2026

Same author

Virion display reveals MD-1 as an endogenous agonist for the orphan receptor GPRC5B.

Science signaling·2026

Same author

Machine Learning-Assisted Rapid Optical Imaging for Label-Free CAR T-Cell Detection in Whole Blood.

Biosensors·2026

Same author

CRISPR-Cas12a2-Based Multiplexed Diagnostic for Rapid and Highly Sensitive Detection of Respiratory Viruses.

Analytical chemistry·2026

Same author

Evaluating Antibody Quality via Simultaneous Size and Charge Measurement with Single Protein Oscillators.

Analytical chemistry·2026

Same author

Hemochromatosis osteoarthritis.

Frontiers in endocrinology·2026

Search research articles

Related Experiment Video

Updated: Dec 11, 2025

Use of Label-free Optical Biosensors to Detect Modulation of Potassium Channels by G-protein Coupled Receptors

Use of Label-free Optical Biosensors to Detect Modulation of Potassium Channels by G-protein Coupled Receptors

Published on: February 10, 2014

Charge-Sensitive Optical Detection of Small Molecule Binding Kinetics in Normal Ionic Strength Buffer.

Runli Liang, Guangzhong Ma, Wenwen Jing

|August 27, 2020

Summary

This summary is machine-generated.

This study introduces a charge-sensitive optical detection (CSOD) technique for accurately measuring small molecule binding kinetics in standard buffers. The new method overcomes limitations of previous techniques, enabling more reliable molecular interaction analysis.

Keywords:

G-protein-coupled receptors binding kinetics charge sensitive optical detection label-free normal ionic strength buffer small molecule virion display

More Related Videos

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

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

Published on: February 18, 2014

Capturing the Interaction Kinetics of an Ion Channel Protein with Small Molecules by the Bio-layer Interferometry Assay

Capturing the Interaction Kinetics of an Ion Channel Protein with Small Molecules by the Bio-layer Interferometry Assay

Published on: March 7, 2018

Related Experiment Videos

Last Updated: Dec 11, 2025

Use of Label-free Optical Biosensors to Detect Modulation of Potassium Channels by G-protein Coupled Receptors

Use of Label-free Optical Biosensors to Detect Modulation of Potassium Channels by G-protein Coupled Receptors

Published on: February 10, 2014

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

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

Published on: February 18, 2014

Capturing the Interaction Kinetics of an Ion Channel Protein with Small Molecules by the Bio-layer Interferometry Assay

Capturing the Interaction Kinetics of an Ion Channel Protein with Small Molecules by the Bio-layer Interferometry Assay

Published on: March 7, 2018

Area of Science:

Biotechnology
Analytical Chemistry
Biosensing

Background:

Label-free detection methods often struggle with small molecules due to mass-based sensitivity limitations.
Existing charge-sensitive optical detection (CSOD) requires diluted buffers, potentially altering molecular binding kinetics.
Ionic strength significantly impacts molecular charge, posing challenges for sensitive detection in physiological conditions.

Purpose of the Study:

To develop an improved CSOD technique for detecting molecular binding kinetics in normal ionic strength buffers.
To overcome the signal loss caused by ionic screening in standard buffer solutions.
To enable accurate kinetic measurements of small molecule interactions with targets like GPCRs.

Main Methods:

An H-shaped sample well was designed to enhance current density at the sensing area, compensating for ionic screening.
Agarose gels were employed to cover electrodes, preventing bubbles from interfering with the sensing area.
The binding kinetics of G-protein-coupled receptors (GPCRs) with small molecule ligands were measured using the developed CSOD system in normal buffer.

Main Results:

The novel CSOD technique successfully measured molecular binding kinetics in normal ionic strength buffers.
Binding affinities measured in normal buffer were found to be stronger compared to those in diluted buffers.
The observed differences in affinity are attributed to electrostatic repulsion effects influenced by buffer ionic strength.

Conclusions:

The developed CSOD technique with an H-shaped well and agarose gel is effective for analyzing molecular binding kinetics in normal buffers.
This advancement allows for more physiologically relevant measurements of small molecule interactions.
The findings highlight the importance of buffer ionic strength in accurately determining molecular binding affinities.