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In Vitro Analysis of E3 Ubiquitin Ligase Function
JoVE Journal
Biology
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JoVE Journal Biology
In Vitro Analysis of E3 Ubiquitin Ligase Function
DOI:

06:06 min

May 14, 2021

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Chapters

  • 00:05Introduction
  • 00:43In Vitro Auto-Ubiquitylation Assay
  • 01:31In vitro Substrate Ubiquitylation Assay
  • 02:13Lysine Discharge Assay
  • 03:37Results: Analysis of the Ubiquitylation Products
  • 05:28Conclusion

Summary

Automatic Translation

The present study provides detailed in vitro ubiquitylation assay protocols for the analysis of E3 ubiquitin ligase catalytic activity. Recombinant proteins were expressed using prokaryotic systems such as Escherichia coli culture.

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