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In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity
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Journal JoVE Biologie
In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity

In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity

DOI:
10.3791/56629-v

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09:45 min

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January 29, 2018

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, , ,
1Institute of Microbiology and Immunology,National Yang-Ming University, 2UC Davis Cancer Center,University of California, Davis, 3Department of Biochemistry and Molecular Medicine,University of California, Davis, 4Institute for Translational Medicine, College of Medical Science and Technology,Taipei Medical University, 5Division of Molecular and Genomic Medicine,National Health Research Institutes, 6Center for Infectious Disease and Cancer Research,Kaohsiung Medical University

Chapitres

  • 00:05Titre
  • 00:39Purification of K-bZIP
  • 03:26SUMOylation Assay
  • 07:15Results: Use of In Vitro SUMOylation Assay to Study the Role of SUMO E3 Ligase in p53 SUMOylation Using Optimal Ubc9 Enzyme Concentration
  • 08:49Conclusion

Summary

Traduction automatique

Traduction automatique

Unlike ubiquitin ligases, few E3 SUMO ligases have been identified. This modified in vitro SUMOylation protocol is able to identify novel SUMO E3 ligases by an in vitro reconstitution assay.

Tags

In Vitro SUMOylationSUMO E3 Ligase ActivitySUMO E3 Ligase IdentificationSUMO Isoform SpecificityProtein PurificationMagnetic Bead PulldownSDS-PAGEProtein QuantificationImage J

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