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Fractionation for Resolution of Soluble and Insoluble Huntingtin Species
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JoVE Journal Biochimica
Fractionation for Resolution of Soluble and Insoluble Huntingtin Species

Fractionation for Resolution of Soluble and Insoluble Huntingtin Species

DOI:
10.3791/57082-v

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07:08 min

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February 27, 2018

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, , ,
1Department of Psychiatry and Human Behavior,University of California Irvine, 2UCI MIND,University of California Irvine, 3Department of Neurobiology and Behavior,University of California Irvine, 4Board of Governors Regenerative Medicine Institute,Cedars-Sinai Medical Center

Capitoli

  • 00:04Titolo
  • 00:51Soluble/Insoluble Fractionation Protocol
  • 03:58Western Blot and Quantification
  • 04:31Results: Representative Results for Soluble and Insoluble Fractionations and Analyses
  • 06:18Conclusion

Summary

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A method is described for fractionation of insoluble and soluble mutant huntingtin species from mouse brain and cell culture. The method described is useful for characterization and quantification of huntingtin protein flux and aids in analyzing protein homeostasis in disease pathogenesis and in the presence of perturbations

Tags

FractionationHuntingtinSolubleInsolubleProtein HomeostasisHuntington’s DiseaseNeurodegenerative DiseaseProtein MisfoldingLysis BufferProtease InhibitorsDounce HomogenizationCell LysateCentrifugationSupernatantPellet

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