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Purification and Quality Control of Recombinant Septin Complexes for Cell-Free Reconstitution
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Purification and Quality Control of Recombinant Septin Complexes for Cell-Free Reconstitution

Purification and Quality Control of Recombinant Septin Complexes for Cell-Free Reconstitution

DOI:
10.3791/63871-v

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11:50 min

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June 23, 2022

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, , , , , ,
1Department of Bionanoscience, Kavli Institute of Nanoscience Delft,Delft University of Technology, 2Institut Fresnel, CNRS UMR7249,Aix Marseille Univ, Centrale Marseille, 3Institut Curie, Université PSL,Sorbonne Université

Chapters

  • 00:05Introduction
  • 00:47Purification of Septin Hetero-Oligomers
  • 04:29Quality Control of Purity and Integrity of the Septin Hetero-Oligomer
  • 07:34Septin Functional Quality Control via Polymerization Analysis
  • 08:57Results: Analysis of Purified Septin Quality
  • 11:01Conclusion

Summary

Automatic Translation

Automatic Translation

In vitro reconstitution of cytoskeletal proteins is a vital tool to understand the basic functional properties of these proteins. The present paper describes a protocol to purify and assess the quality of recombinant septin complexes, which play a central role in cell division and migration.

Tags

Keywords: Recombinant Septin ComplexesCell-free ReconstitutionPurificationQuality ControlAffinity ChromatographyHis-tagged SeptinsStrep-II Tagged ProteinsFPLCBacterial ExpressionIPTG InductionCell LysisSonicationCentrifugationOptical DensityMsfGFPMEGFPProtein Degradation

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