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Efficient and Scalable Production of Full-length Human Huntingtin Variants in Mammalian Cells using a Transient Expression System
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Efficient and Scalable Production of Full-length Human Huntingtin Variants in Mammalian Cells using a Transient Expression System

Efficient and Scalable Production of Full-length Human Huntingtin Variants in Mammalian Cells using a Transient Expression System

DOI:
10.3791/63190-v

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10:52 min

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December 10, 2021

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, , , , , ,
1Discovery Biology,Curia Global Inc.

Chapters

  • 00:05Introduction
  • 00:24Large-Scale Transfection of HEK293 Cells by Polyethyleneimine
  • 03:10Fast Protein Liquid Chromatography (FPLC) Purification of HTT Using Anti-FLAG Column and SEC
  • 05:56Analytical HPLC SEC-MALS-dRI to Analyze HTT Polydispersity
  • 07:49Results: Qualitative and Quantitative Estimation of Purified FL HTT
  • 10:12Conclusion

Summary

Automatic Translation

Automatic Translation

We provide scalable protocols covering construct design, transient transfection, and expression and purification of full-length human huntingtin protein variants in HEK293 cells.

Tags

Keywords: HuntingtinTransient ExpressionHEK293 CellsPolyethyleneimineTransfectionSodium ButyrateAnti-FLAG PurificationFPLCProtein ProductionHuntington’s Disease

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