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Related Concept Videos

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...
Protein-protein Interfaces02:04

Protein-protein Interfaces

Many proteins form complexes to carry out their functions, making protein-protein interactions (PPIs) essential for an organism's survival. Most PPIs are stabilized by numerous weak noncovalent chemical forces. The physical shape of the interfaces determines the way two proteins interact. Many globular proteins have closely-matching shapes on their surfaces, which form a large number of weak bonds. Additionally, many PPIs occur between two helices or between a surface cleft and a polypeptide...

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Related Experiment Video

Updated: Jul 19, 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 interaction screening by quantitative immunoprecipitation combined with knockdown (QUICK).

Matthias Selbach1, Matthias Mann

  • 1Max Planck Institute of Biochemistry, Department of Proteomics and Signal Transduction, Am Klopferspitz 18, D-82152 Martinsried, Germany.

Nature Methods
|October 31, 2006
PubMed
Summary

This study introduces a new method to identify protein interactions in cells using stable isotope labeling with amino acids in cell culture (SILAC) and other techniques. This approach enhances the accuracy of detecting endogenous protein-protein interactions (PPIs).

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Identification of Protein Interaction Partners in Mammalian Cells Using SILAC-immunoprecipitation Quantitative Proteomics
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Identification of Protein Interaction Partners in Mammalian Cells Using SILAC-immunoprecipitation Quantitative Proteomics

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Related Experiment Videos

Last Updated: Jul 19, 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

Quantification of Protein Interaction Network Dynamics using Multiplexed Co-Immunoprecipitation
07:57

Quantification of Protein Interaction Network Dynamics using Multiplexed Co-Immunoprecipitation

Published on: August 21, 2019

Identification of Protein Interaction Partners in Mammalian Cells Using SILAC-immunoprecipitation Quantitative Proteomics
12:53

Identification of Protein Interaction Partners in Mammalian Cells Using SILAC-immunoprecipitation Quantitative Proteomics

Published on: July 6, 2014

Area of Science:

  • Cellular Biology
  • Biochemistry
  • Proteomics

Background:

  • Current protein-protein interaction (PPI) screening methods often use overexpressed artificial proteins, limiting their relevance to in vivo conditions.
  • Assessing interactions of endogenous proteins is crucial for understanding cellular mechanisms.

Purpose of the Study:

  • To develop and validate a high-confidence method for detecting endogenous protein-protein interactions (PPIs) in mammalian cells.
  • To identify novel interaction partners for key cellular proteins like beta-catenin and Cbl.

Main Methods:

  • Integration of stable isotope labeling with amino acids in cell culture (SILAC) for quantitative analysis.
  • Application of RNA interference (RNAi) for target protein knockdown.
  • Utilizing coimmunoprecipitation to isolate protein complexes.
  • Employing quantitative mass-spectrometry for sensitive detection and identification of interaction partners.

Main Results:

  • The combined approach successfully identified cellular interaction partners of endogenous proteins with high confidence.
  • Specific interaction partners for beta-catenin and Cbl were successfully identified using this novel screening strategy.

Conclusions:

  • This integrated proteomic approach provides a robust platform for studying endogenous protein-protein interactions (PPIs) in their native cellular context.
  • The method significantly improves the reliability and confidence in identifying in vivo protein interaction networks.