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Sequential Extraction of Soluble and Insoluble Alpha-Synuclein from Parkinsonian Brains
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Sequential Extraction of Soluble and Insoluble Alpha-Synuclein from Parkinsonian Brains

Sequential Extraction of Soluble and Insoluble Alpha-Synuclein from Parkinsonian Brains

DOI:
10.3791/53415-v

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09:27 min

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January 05, 2016

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1Reta Lila Weston Institute of Neurological Studies,UCL Institute of Neurology

Chapters

  • 00:05Title
  • 00:36Brain Sample Homogenization and Differential Ultracentrifugation
  • 04:37Immunoblotting of Samples
  • 07:25Results: Distinct Species of Alpha-Synuclein Detected by Western Immunoblot
  • 08:35Conclusion

Summary

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Here, we present a protocol for the isolation of increasingly insoluble/aggregated alpha-synuclein (α-syn) from post-mortem human brain tissue. Through the utilization of buffers with increasing detergent strength and high-speed ultracentrifugation techniques, the variable properties of α-syn aggregation in diseased and non-diseased tissue can be examined.

Tags

Keywords: Alpha-synucleinParkinson’s DiseaseSequential ExtractionSolubleInsolubleBasal GangliaHomogenizationUltracentrifugationTBSSDSUrea

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