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Related Experiment Video

Updated: Dec 30, 2025

Purification of Ubiquitinated p53 Proteins from Mammalian Cells
10:55

Purification of Ubiquitinated p53 Proteins from Mammalian Cells

Published on: March 21, 2022

2.6K

Interaction of p53 with cellular proteins.

David M Keller1, Shelya X Zeng, Hua Lu

  • 1Department of Biochemistry and Molecular Biology, Oregon Health and Science University, Portland, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 26, 2003
PubMed
Summary

Identifying p53-binding proteins is crucial for understanding its tumor suppressor functions. This chapter details glutathione-S-transferase (GST) and co-immunoprecipitation (co-IP) assays for characterizing these interactions.

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Area of Science:

  • Molecular Biology
  • Cancer Research
  • Cellular Signaling

Background:

  • p53 is a critical tumor suppressor protein.
  • Cellular proteins interacting with p53 modulate its regulatory functions.
  • Complete understanding of p53 biology requires characterization of its interacting partners.

Purpose of the Study:

  • To outline methods for identifying and characterizing p53-binding proteins.
  • To provide practical guidance for implementing protein-protein interaction assays.
  • To facilitate comprehensive studies of p53-mediated cellular responses.

Main Methods:

  • Glutathione-S-transferase (GST) protein-protein interaction assay.
  • Co-immunoprecipitation (co-IP) assay.
  • Detailed protocols and optimization tips for both assays.

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Last Updated: Dec 30, 2025

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Yeast As a Chassis for Developing Functional Assays to Study Human P53
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Main Results:

  • The described methods enable reliable identification of p53-interacting proteins.
  • GST and co-IP assays are shown to be cost-efficient and reproducible.
  • The techniques are accessible for laboratories without specialized expertise.

Conclusions:

  • Characterizing p53-interacting proteins is essential for a complete understanding of p53 tumor suppressor activity.
  • GST and co-IP assays offer practical, efficient, and reliable approaches for this characterization.
  • These methods can be readily adopted to advance p53 research.