Journal
/
Biology
/
Method for Measuring the Activity of Deubiquitinating Enzymes in Cell Lines and Tissue Samples
JoVE Journal
Biology
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biology
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

•

09:45 min

•

May 10, 2015

•

, , , , ,
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

Chapters

  • 00:05Title
  • 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

Automatic Translation

Automatic Translation

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

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

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/52784?article_type=v

Actin Co-Sedimentation Assay; for the Analysis of Protein Binding to F-Actin
check_url/52784?article_type=v

Quantitation of γH2AX Foci in Tissue Samples
check_url/52784?article_type=v

An Assay for Measuring the Activity of Escherichia coli Inducible Lysine Decarboxyase
check_url/52784?article_type=v

Do-It-Yourself Device for Recovery of Cryopreserved Samples Accidentally Dropped into Cryogenic Storage Tanks
check_url/52784?article_type=v

Live Imaging of Apoptotic Cell Clearance during Drosophila Embryogenesis
check_url/52784?article_type=v

Measuring In Vitro ATPase Activity for Enzymatic Characterization
check_url/52784?article_type=v

Comparison of Three Different Methods for Determining Cell Proliferation in Breast Cancer Cell Lines
check_url/52784?article_type=v

Measuring Lactase Enzymatic Activity in the Teaching Lab
check_url/52784?article_type=v

Hybrid-Cut: An Improved Sectioning Method for Recalcitrant Plant Tissue Samples
check_url/52784?article_type=v

Dissection of the Auditory Bulla in Postnatal Mice: Isolation of the Middle Ear Bones and Histological Analysis

Read Article