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

Immunoprecipitation

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...

Western Blotting

Western Blotting

Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.

You might also read

Related Articles

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

Sort by

Same author

LZTS2 Emerges as a Regulator of Craniofacial Development and Modulator of DYRK1A.

bioRxiv : the preprint server for biology·2026

Same author

Glycan Profiling Identifies Chondroitin-4-sulfate as a Biomarker for Platinum Response and Therapeutic Target in Ovarian Cancer.

bioRxiv : the preprint server for biology·2025

Same author

Proteasome inhibition paradoxically degrades gain-of-function mutant p53 R273H in NSCLC and could have therapeutic implications.

Frontiers in oncology·2024

Same author

Dyrk1a is required for craniofacial development in Xenopus laevis.

Developmental biology·2024

Same author

Dyrk1a is required for craniofacial development in <i>Xenopus laevis</i>.

bioRxiv : the preprint server for biology·2024

Same author

Insights from the protein interaction Universe of the multifunctional "Goldilocks" kinase DYRK1A.

Frontiers in cell and developmental biology·2023

Search research articles

Related Experiment Video

Updated: Dec 20, 2025

Protein Extract Preparation and Co-immunoprecipitation from Caenorhabditis elegans

Protein Extract Preparation and Co-immunoprecipitation from Caenorhabditis elegans

Published on: May 23, 2020

Preparing Immunoprecipitations for Immunoblotting.

Larisa Litovchick

Cold Spring Harbor Protocols

|June 3, 2020

Summary

This study details efficient methods for eluting immunoprecipitated proteins from beads for immunoblotting analysis. Boiling in SDS-PAGE sample buffer is a cost-effective method for protein elution and subsequent gel electrophoresis.

Area of Science:

Biochemistry
Molecular Biology
Immunology

Background:

Immunoprecipitation (IP) is a common technique for isolating specific proteins.
Analysis of immunoprecipitated proteins often involves immunoblotting.
Efficient elution of proteins from affinity beads is crucial for downstream analysis.

Purpose of the Study:

To describe and compare methods for eluting immunoprecipitated proteins from affinity beads.
To provide a protocol for preparing samples for protein gel electrophoresis after immunoprecipitation.

Main Methods:

Elution using SDS-PAGE sample loading buffer and heat (95°C).
Elution using low (pH 2.1-2.8) or high (pH 10-11) pH conditions.
Elution using competing peptides (where applicable).

More Related Videos

Author Spotlight: Unraveling the Molecular Mechanisms of Brown and Beige Adipocyte Regulation

Author Spotlight: Unraveling the Molecular Mechanisms of Brown and Beige Adipocyte Regulation

Published on: January 5, 2024

Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling

Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling

Published on: November 17, 2019

Related Experiment Videos

Last Updated: Dec 20, 2025

Protein Extract Preparation and Co-immunoprecipitation from Caenorhabditis elegans

Protein Extract Preparation and Co-immunoprecipitation from Caenorhabditis elegans

Published on: May 23, 2020

Author Spotlight: Unraveling the Molecular Mechanisms of Brown and Beige Adipocyte Regulation

Author Spotlight: Unraveling the Molecular Mechanisms of Brown and Beige Adipocyte Regulation

Published on: January 5, 2024

Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling

Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling

Published on: November 17, 2019

Main Results:

SDS-PAGE sample buffer with boiling effectively elutes proteins and prepares them for electrophoresis.
Low or high pH elution can also release proteins but may affect antibody integrity.
Competing peptide elution removes antibodies but is not always feasible or cost-effective.

Conclusions:

Boiling in SDS-PAGE sample buffer is a practical and efficient method for eluting immunoprecipitated proteins for immunoblotting.
The choice of elution method depends on downstream application requirements and cost considerations.
This protocol facilitates routine immunoprecipitation and immunoblotting experiments.