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生物化学
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Generation of Native, Untagged Huntingtin Exon1 Monomer and Fibrils Using a SUMO Fusion Strategy
JoVE 杂志
生物化学
This content is Free Access.
JoVE 杂志 生物化学
Generation of Native, Untagged Huntingtin Exon1 Monomer and Fibrils Using a SUMO Fusion Strategy

Generation of Native, Untagged Huntingtin Exon1 Monomer and Fibrils Using a SUMO Fusion Strategy

DOI:
10.3791/57506-v

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11:22 min

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June 27, 2018

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, , ,
1Laboratory of Molecular and Chemical Biology of Neurodegeneration,Ecole Polytechnique Fédérale de Lausanne (EPFL)

Chapters

  • 00:04Title
  • 00:53Cell Lysis and Purification of His6-SUMO Httex1 Fusion Protein by Immobilized Metal Affinity Chromatography (IMAC)
  • 02:56Cleavage of the His6-SUMO-tag and HPLC Purification
  • 06:06Disaggregation and Resolubilization of Httex1 Proteins
  • 07:53Results: Characterization of Native, Untagged Huntingtin Exon1 Monomer and Fibrils Prepared via SUMO Fusion
  • 09:52Conclusion

Summary

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Here, we present a robust and optimized protocol for the production of milligram quantities of native, tag-free monomers and fibrils of the exon1 of the Huntingtin protein (Httex1) based on the transient fusion of small ubiquitin related modifier (SUMO).

Tags

Keywords: Native Huntingtin Exon1SUMO Fusion StrategySDS-PAGE AnalysisIMAC PurificationHPLC PurificationDTTL-cysteineTrifluoroacetic AcidAggregation-prone ProteinsAmyloid-betaAlpha-synuclein

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