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

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

1,665 Views

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11:50 min

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June 23, 2022

DOI:

10.3791/63871-v

DOI:
10.3791/63871-v

•

11:50 min
June 23, 2022

•

7 Views
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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

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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.

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

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Castro-Linares, G., den Haan, J., Iv, F., Silva Martins, C., Bertin, A., Mavrakis, M., Koenderink, G. H. Purification and Quality Control of Recombinant Septin Complexes for Cell-Free Reconstitution. J. Vis. Exp. (184), e63871, doi:10.3791/63871 (2022).

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