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Quantifying Subcellular Ubiquitin-proteasome Activity in the Rodent Brain
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Quantifying Subcellular Ubiquitin-proteasome Activity in the Rodent Brain
DOI:

09:25 min

May 21, 2019

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Chapters

  • 00:04Title
  • 00:44Nuclear and Cytoplasmic Extraction
  • 02:31Synaptic Fraction Collection
  • 03:45Proteasome Activity Assay
  • 06:36Quantification of Linkage-specific Protein Ubiquitination
  • 07:34Results: Differential Subcellular Changes in Proteasome Activity and Protein Polyubiquitination in the Rat Amygdala Following Learning
  • 09:13Conclusion

Summary

Automatic Translation

This protocol is designed to efficiently quantify ubiquitin-proteasome system (UPS) activity in different cellular compartments of the rodent brain. Users are able to examine UPS functioning in nuclear, cytoplasmic and synaptic fractions in the same animal, reducing the amount of time and number of animals needed to perform these complex analyses.

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