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

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...
Immunoprecipitation01:20

Immunoprecipitation

Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...

You might also read

Related Articles

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

Sort by
Same author

Localization of Organelle Proteins Using Data-Independent Acquisition (DIA-LOP).

Molecular & cellular proteomics : MCPĀ·2025
Same author

A guide to the clinical management of attrition.

British dental journalĀ·2018
Same author

Investigation of the erosive potential of sour novelty sweets.

British dental journalĀ·2017
Same author

The availability of novelty sweets within high school localities.

British dental journalĀ·2016
Same author

A cross-sectional study of dentine sensitivity in periodontitis patients in Trinidad and Tobago.

International journal of dental hygieneĀ·2016
Same author

Impact of Azithromycin on the Quorum Sensing-Controlled Proteome of Pseudomonas aeruginosa.

PloS oneĀ·2016

Related Experiment Video

Updated: May 31, 2026

A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation
14:44

A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation

Published on: September 24, 2012

Method for suppressing non-specific protein interactions observed with affinity resins.

J S Rees1, K S Lilley

  • 1Cambridge Centre for Proteomics, Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK. jsr31@cam.ac.uk

Methods (San Diego, Calif.)
|July 5, 2011
PubMed
Summary
This summary is machine-generated.

Researchers adapted a method using thiocyanate anions to reduce unwanted proteins in affinity purification. This technique helps enrich specific protein interactions, though its effectiveness varies with the bait protein's natural abundance.

More Related Videos

Protein Complex Affinity Capture from Cryomilled Mammalian Cells
10:37

Protein Complex Affinity Capture from Cryomilled Mammalian Cells

Published on: December 9, 2016

Pulldown Assay Coupled with Co-Expression in Bacteria Cells as a Time-Efficient Tool for Testing Challenging Protein-Protein Interactions
07:03

Pulldown Assay Coupled with Co-Expression in Bacteria Cells as a Time-Efficient Tool for Testing Challenging Protein-Protein Interactions

Published on: December 23, 2022

Related Experiment Videos

Last Updated: May 31, 2026

A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation
14:44

A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation

Published on: September 24, 2012

Protein Complex Affinity Capture from Cryomilled Mammalian Cells
10:37

Protein Complex Affinity Capture from Cryomilled Mammalian Cells

Published on: December 9, 2016

Pulldown Assay Coupled with Co-Expression in Bacteria Cells as a Time-Efficient Tool for Testing Challenging Protein-Protein Interactions
07:03

Pulldown Assay Coupled with Co-Expression in Bacteria Cells as a Time-Efficient Tool for Testing Challenging Protein-Protein Interactions

Published on: December 23, 2022

Area of Science:

  • Biochemistry
  • Proteomics
  • Molecular Biology

Background:

  • High-throughput protein complex purification often yields contaminating proteins.
  • These contaminants can be specific to the affinity matrix or introduced tags.
  • Non-specific binding complicates the identification of true protein interactors.

Purpose of the Study:

  • To adapt and evaluate the use of thiocyanate anions for reducing common contaminants in affinity purification.
  • To assess the impact of this method on the enrichment of specific protein binding partners.
  • To investigate the influence of bait protein abundance on the efficacy of contaminant reduction.

Main Methods:

  • Adaptation of pre-equilibration of affinity surfaces with thiocyanate anions.
  • Application of the method in affinity purification experiments.
  • Analysis of protein complex composition to identify contaminants and specific partners.

Main Results:

  • The thiocyanate anion pre-equilibration method was successfully adapted.
  • The method demonstrated a reduction in common contaminating proteins.
  • The effectiveness of contaminant reduction varied depending on the specific bait protein used, likely linked to its endogenous abundance.

Conclusions:

  • Thiocyanate anion pre-equilibration is a viable strategy for minimizing non-specific protein binding in affinity purification.
  • The method shows promise for improving the quality of proteomic data by enriching specific interactions.
  • Further studies are needed to optimize the application based on bait protein characteristics.