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

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Atomic Force Microscopy01:08

Atomic Force Microscopy

Atomic force microscopy (AFM) is a type of scanning probe microscopy that can analyze topographic details of various specimens like ceramics, glass, polymers, and biological samples. AFM offers over 1000 times more resolution than the optical imaging system. Images generated from AFM are three-dimensional surface profiles, offering an advantage over the flat, two-dimensional images from other imaging techniques.
The AFM Probe
The probe is regarded as the heart of any AFM setup and comprises the...

You might also read

Related Articles

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

Sort by
Same author

Patient Safety Issues From Information Overload in Electronic Medical Records.

Journal of patient safety·2022
Same author

A quantitative LC-MS/MS approach for monitoring 2'-fluoro-2'-deoxy-D-glucose uptake in tumor tissue.

Bioanalysis·2021
Same author

A Flexible Multiplatform Bioanalytical Strategy for Measurement of Total Circulating Shed Target Receptors: Application to Soluble B Cell Maturation Antigen Levels in the Presence of a Bispecific Antibody Drug.

Assay and drug development technologies·2020
Same author

Considerations for Soluble Protein Biomarker Blood Sample Matrix Selection.

The AAPS journal·2020
Same author

Characterization of the Selective Indoleamine 2,3-Dioxygenase-1 (IDO1) Catalytic Inhibitor EOS200271/PF-06840003 Supports IDO1 as a Critical Resistance Mechanism to PD-(L)1 Blockade Therapy.

Molecular cancer therapeutics·2018
Same author

Workshop Report: Crystal City VI-Bioanalytical Method Validation for Biomarkers.

The AAPS journal·2016

Related Experiment Video

Updated: Jun 23, 2026

Force Spectroscopy of Single Protein Molecules Using an Atomic Force Microscope
06:45

Force Spectroscopy of Single Protein Molecules Using an Atomic Force Microscope

Published on: February 28, 2019

Association kinetics from single molecule force spectroscopy measurements.

Senli Guo1, Nimit Lad, Chad Ray

  • 1Department of Chemistry, Duke University, Durham, North Carolina 27708, USA.

Biophysical Journal
|April 23, 2009
PubMed
Summary

We developed a new atomic force microscopy method to measure molecular association energy. This technique quantifies the activation energy for binding, offering insights into molecular interactions.

More Related Videos

Investigating Single Molecule Adhesion by Atomic Force Spectroscopy
09:48

Investigating Single Molecule Adhesion by Atomic Force Spectroscopy

Published on: February 27, 2015

Covalent Attachment of Single Molecules for AFM-based Force Spectroscopy
10:37

Covalent Attachment of Single Molecules for AFM-based Force Spectroscopy

Published on: March 16, 2020

Related Experiment Videos

Last Updated: Jun 23, 2026

Force Spectroscopy of Single Protein Molecules Using an Atomic Force Microscope
06:45

Force Spectroscopy of Single Protein Molecules Using an Atomic Force Microscope

Published on: February 28, 2019

Investigating Single Molecule Adhesion by Atomic Force Spectroscopy
09:48

Investigating Single Molecule Adhesion by Atomic Force Spectroscopy

Published on: February 27, 2015

Covalent Attachment of Single Molecules for AFM-based Force Spectroscopy
10:37

Covalent Attachment of Single Molecules for AFM-based Force Spectroscopy

Published on: March 16, 2020

Area of Science:

  • Biophysics
  • Chemical Physics
  • Materials Science

Background:

  • Single molecule force spectroscopy typically measures dissociation kinetics.
  • Understanding association kinetics is crucial for molecular interactions.
  • Existing methods for measuring association energy are limited.

Purpose of the Study:

  • To present a novel atomic force microscopy (AFM)-based approach to determine the activation energy of association reactions.
  • To approximate the grafting density of reactive receptors.
  • To provide a method compatible with existing force spectroscopy setups.

Main Methods:

  • Utilizing AFM to apply mechanical force and measure bond formation probability.
  • Analyzing the dependence of bond formation probability on probe velocity.
  • Tethering one molecule to the AFM probe via a flexible polymeric linker.
  • Considering and correcting for potential errors in activation energy measurements.

Main Results:

  • Successfully measured the activation energy of biotin-streptavidin association.
  • Obtained an activation energy value of 8 ± 1 kT, including steric factor contributions.
  • The measured value aligns with previously reported solution-based association rates.

Conclusions:

  • The developed AFM approach provides a reliable method for measuring association activation energy.
  • The kinetics of biotin-streptavidin binding are primarily governed by the reaction step.
  • This method enhances the study of molecular interactions using force spectroscopy.