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

Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and reduce chemical noise during analyte detection. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called...
Affinity Chromatography01:03

Affinity Chromatography

Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...
Peptide Identification Using Tandem Mass Spectrometry01:33

Peptide Identification Using Tandem Mass Spectrometry

Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
MALDI-TOF Mass Spectrometry01:19

MALDI-TOF Mass Spectrometry

Mass spectrometry is a powerful characterization technique that can identify and separate a wide variety of compounds ranging from chemical to biological entities, based on their mass-to-charge ratio (m/z). The instruments that allow this detection, known as mass spectrometers, have three components: an ion source, a mass analyzer, and a detector. These spectrometers differ based on the nature of their ion source and analyzers.Matrix-assisted laser desorption ionization (MALDI) is a commonly...
Mass Spectrometry: Overview01:19

Mass Spectrometry: Overview

Mass spectrometry is an analytical technique used to determine the molecular mass and molecular formula of a compound. The basic principle of mass spectrometry is to generate ions from the analyte molecule and measure these ion abundances against their molecular mass. One common type of ionization, known as electron ionization or EI, bombards the analyte molecules in the gas phase with high-energy electron beams. The electron beams displace an electron from the molecule and leave behind a...

You might also read

Related Articles

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

Sort by
Same author

SARS-CoV-2 infection in female sex workers from Nairobi, Kenya early in the COVID-19 pandemic: Seroincidence and behavioural associations.

PloS one·2026
Same author

Findings from comprehensive genome sequencing in the Canadian population: Results from the GENCOV Study.

Genetics in medicine open·2026
Same author

SCM-1/SCAMP Maintains Microdomain Boundaries and Cargo Sorting within the Endosomal System.

bioRxiv : the preprint server for biology·2026
Same author

Serological Benefit of SARS-CoV-2 Vaccination Relative to Infection in Children With Acute Lymphoblastic Leukemia.

Pediatric blood & cancer·2026
Same author

The molecular glue CLEO4-88 inhibits the ACAA1 thiolase by induced binding to GID4.

Nature chemical biology·2026
Same author

3in1: an ultra-fast phage display biopanning method overcoming the biopanning bias and yielding more diverse binders.

BioTechniques·2026

Related Experiment Video

Updated: May 22, 2026

Analyzing Protein Architectures and Protein-Ligand Complexes by Integrative Structural Mass Spectrometry
07:33

Analyzing Protein Architectures and Protein-Ligand Complexes by Integrative Structural Mass Spectrometry

Published on: October 15, 2018

Affinity-purification coupled to mass spectrometry: basic principles and strategies.

Wade H Dunham1, Michael Mullin, Anne-Claude Gingras

  • 1Samuel Lunenfeld Research Institute at Mount Sinai Hospital, Toronto, ON, Canada.

Proteomics
|May 22, 2012
PubMed
Summary

Understanding protein function requires identifying protein-protein interactions. Affinity-purification followed by mass spectrometry (AP-MS) is a key technique for discovering these interactions, aided by biochemical and computational tools.

More Related Videos

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling
09:35

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

Published on: April 1, 2017

Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization
12:11

Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization

Published on: February 27, 2020

Related Experiment Videos

Last Updated: May 22, 2026

Analyzing Protein Architectures and Protein-Ligand Complexes by Integrative Structural Mass Spectrometry
07:33

Analyzing Protein Architectures and Protein-Ligand Complexes by Integrative Structural Mass Spectrometry

Published on: October 15, 2018

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling
09:35

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

Published on: April 1, 2017

Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization
12:11

Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization

Published on: February 27, 2020

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Proteomics

Background:

  • Protein function is often elucidated by studying its interactions with other proteins.
  • Identifying interactions with known proteins is crucial for characterizing unknown proteins.
  • Affinity-purification followed by mass spectrometry (AP-MS) has emerged as a powerful method for studying protein-protein interactions.

Purpose of the Study:

  • To review the fundamental principles of affinity-purification coupled to mass spectrometry (AP-MS).
  • To highlight the essential biochemical and computational tools used in AP-MS for protein interaction discovery.
  • To emphasize the reporting of high-quality protein-protein interaction data.

Main Methods:

  • Review of affinity-purification techniques.
  • Overview of mass spectrometry principles for protein identification.
  • Discussion of computational tools for analyzing AP-MS data.

Main Results:

  • AP-MS is a robust method for identifying and quantifying protein-protein interactions.
  • Biochemical and computational tools enhance the discovery and validation of protein interactions.
  • The review emphasizes strategies for obtaining high-quality interaction data.

Conclusions:

  • AP-MS is a vital technique in modern proteomics for understanding protein function.
  • Effective use of AP-MS relies on both optimized biochemical protocols and sophisticated computational analysis.
  • High-quality protein-protein interaction data is essential for advancing biological research.