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

You might also read

Related Articles

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

Sort by

Same author

Differences in canine serum N-glycosylation pattern during infection and pregnancy.

Scientific reports·2026

Same author

Pregnancy-related changes in the canine serum N-glycosylation pattern studied by Rapifluor HILIC-UPLC-FLR-MS.

Scientific reports·2024

Same author

Oxylipin profiling by LC-ESI-MS/MS in canine serum and plasma to investigate ovulation-specific changes.

Prostaglandins & other lipid mediators·2023

Same author

Circulating proteins reveal prior use of menopausal hormonal therapy and increased risk of breast cancer.

Translational oncology·2022

Same author

Systems Biology Approaches to Decipher the Underlying Molecular Mechanisms of Glioblastoma Multiforme.

International journal of molecular sciences·2021

Same author

Next generation plasma proteome profiling of COVID-19 patients with mild to moderate symptoms.

EBioMedicine·2021

Same journal

LaeA Orchestrates Iron-Heme Supply and P450 Catalytic Efficiency for Enhanced Echinocandin B Biosynthesis in Aspergillus nidulans.

Biotechnology journal·2026

Same journal

Emerging Bioengineering Technologies in Female Reproduction: Preclinical Advances, Translational Challenges, and Future Outlook.

Biotechnology journal·2026

Same journal

Multi-Enzyme Cascade Reaction of Crude Enzyme Strategy for the Economical and Efficient Bioconversion of Rebaudioside A to Rebaudioside M.

Biotechnology journal·2026

Same journal

Metabolic and Synthetic Biology Strategies for Enhancing Single-Cell Protein Production in Saccharomyces cerevisiae.

Biotechnology journal·2026

Same journal

Severe Restriction of Glucose Import Enhances Recombinant Protein Production in Escherichia coli.

Biotechnology journal·2026

Same journal

Butyrate-Producing Bacteria in Intestinal Disease Therapy: Potential and Challenges.

Biotechnology journal·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Jun 5, 2026

Protein Complex Affinity Capture from Cryomilled Mammalian Cells

Protein Complex Affinity Capture from Cryomilled Mammalian Cells

Published on: December 9, 2016

Targeted protein pullout from human tissue samples using competitive elution.

Ronny Falk¹, Margareta Ramström, Cecilia Eriksson

¹Royal Institute of Technology, AlbaNova University Center, Department of Biotechnology, Stockholm, Sweden.

Biotechnology Journal

|December 21, 2010

Summary

This summary is machine-generated.

This study introduces a novel competitive elution method using antibodies and recombinant antigens for selective protein pullout. This technique enhances accuracy in identifying target proteins and their interactions, reducing background noise for downstream analysis.

More Related Videos

Identification of Protein Interacting Partners Using Tandem Affinity Purification

Identification of Protein Interacting Partners Using Tandem Affinity Purification

Published on: February 25, 2012

2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes

2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes

Published on: August 6, 2018

Related Experiment Videos

Last Updated: Jun 5, 2026

Protein Complex Affinity Capture from Cryomilled Mammalian Cells

Protein Complex Affinity Capture from Cryomilled Mammalian Cells

Published on: December 9, 2016

Identification of Protein Interacting Partners Using Tandem Affinity Purification

Identification of Protein Interacting Partners Using Tandem Affinity Purification

Published on: February 25, 2012

2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes

2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes

Published on: August 6, 2018

Area of Science:

Proteomics
Biochemistry
Molecular Biology

Background:

Protein isolation is key for cellular information gathering, often using antibodies.
Specificity of antibodies and elution conditions are critical to prevent false positives.
Efficient elution minimizes background proteins, improving downstream analysis.

Purpose of the Study:

To present a novel approach for selective protein pullout using competitive elution.
To enhance the accuracy of identifying target proteins and their interaction partners.
To reduce the effort in downstream analyses by minimizing eluted background proteins.

Main Methods:

Utilized monospecific antibodies for selective protein pullout from complex biosamples.
Employed competitive elution with recombinant antigens to release target proteins.
Used IMAC spin columns to deplete His-tagged antigens from the eluted sample.
Applied the method to a model system (recombinant BBC7) and a native human protein (cTAGE5).

Main Results:

Confirmed affinity isolation of target proteins via SDS-PAGE and mass spectrometry.
Demonstrated more selective elution with the competitive method compared to low pH elution.
Successfully extracted both spiked and endogenous target proteins with high specificity.

Conclusions:

Competitive elution offers an effective strategy for selective protein pullout.
This method improves the accuracy of identifying proteins and their interaction partners.
The technique is suitable for wide-scale pullout experiments in proteomics research.