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Purification of Ubiquitinated p53 Proteins from Mammalian Cells
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JoVE 杂志 生物化学
Purification of Ubiquitinated p53 Proteins from Mammalian Cells
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Purification of Ubiquitinated p53 Proteins from Mammalian Cells

DOI:
10.3791/63602-v

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10:55 min

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March 21, 2022

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, ,
1Center for Translational Medicine,The Affiliated Zhangjiagang Hospital of Soochow University

Chapters

  • 00:05Introduction
  • 00:43Plasmid Transfection
  • 02:16Cell Collection
  • 02:52Purification of Ubiquitinated Proteins under Nondenaturing Conditions
  • 05:01Purification of Ubiquitinated Proteins under Denaturing Conditions
  • 07:49Detection of Purified Ubiquitinated Proteins by Western Blotting
  • 08:29Results: Ubiquitinated p53 Proteins Purified under Nondenaturing and Denaturing Conditions
  • 10:22Conclusion

Summary

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The protocol describes a step-by-step method to purify ubiquitinated proteins from mammalian cells using the p53 tumor suppressor protein as an example. Ubiquitinated p53 proteins were purified from cells under stringent nondenaturing and denaturing conditions.

Tags

Ubiquitinated P53Protein PurificationMammalian CellsUbiquitinationLiposome TransfectionMG132FLAG Lysis BufferProtease Inhibitor

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