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Method for Measuring the Activity of Deubiquitinating Enzymes in Cell Lines and Tissue Samples
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Journal JoVE Biologie
Method for Measuring the Activity of Deubiquitinating Enzymes in Cell Lines and Tissue Samples

Method for Measuring the Activity of Deubiquitinating Enzymes in Cell Lines and Tissue Samples

DOI:
10.3791/52784-v

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09:45 min

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May 10, 2015

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, , , , ,
1Department of Neuroscience,University of Minnesota, 2Institute for Translational Neuroscience,University of Minnesota, 3Department of Obstetrics, Gynecology, and Women’s Heath,University of Minnesota, 4Masonic Cancer Center,University of Minnesota

Chapitres

  • 00:05Titre
  • 01:09Culturing Cells for the Experiment
  • 02:24Cell Harvesting and Cell Lysis
  • 04:40Sample Derivatization
  • 06:03Western Blot
  • 07:42Results: Analysis of Deubiquinating Enzyme Activity
  • 09:07Conclusion

Summary

Traduction automatique

Traduction automatique

The current protocol details a method for measuring the activity of functionally homologous deubiquitinating enzymes. Specialized probes covalently modify the enzyme and allow for detection. This method holds the potential to identify new therapeutic targets.

Tags

Deubiquitinating EnzymesUbiquitin-proteasome SystemCell LinesActivity AssayProteasomal DegradationActive Site ProbesEnzyme ActivityExpressionCell DisruptionPathologiesNeurodegenerative DiseasesCancer

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