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Recombinant Protein Expression for Structural Biology in HEK 293F Suspension Cells: A Novel and Accessible Approach
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Biologie
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Journal JoVE Biologie
Recombinant Protein Expression for Structural Biology in HEK 293F Suspension Cells: A Novel and Accessible Approach

Recombinant Protein Expression for Structural Biology in HEK 293F Suspension Cells: A Novel and Accessible Approach

DOI:
10.3791/51897-v

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11:20 min

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October 16, 2014

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, , , , , , ,
1Department of Biochemistry,University of Leicester

Chapitres

  • 00:05Titre
  • 01:48Starter Cultures
  • 03:43Large-scale Culture/Transfection in 1 L Roller Bottles
  • 05:16Protein-complex Purification from Whole Cell Extract
  • 09:20Results: Expression and Purification of the HDAC1, SDS3, and Sin3A Ternary Complex
  • 10:35Conclusion

Summary

Traduction automatique

Traduction automatique

The expression of recombinant proteins by mammalian systems is becoming an attractive method for producing protein complexes for structural biology. Here we present a simple yet highly efficient expression system using suspension grown mammalian cells to purify protein complexes for structural studies.

Tags

Keywords: Recombinant Protein ExpressionStructural BiologyHEK 293F Suspension CellsEukaryotic ExpressionProtein PurificationPost-translational ModificationsProtein ComplexesTransient TransfectionMammalian Cell Lines

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