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Efficient Mammalian Cell Expression and Single-step Purification of Extracellular Glycoproteins for Crystallization
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Efficient Mammalian Cell Expression and Single-step Purification of Extracellular Glycoproteins for Crystallization

Efficient Mammalian Cell Expression and Single-step Purification of Extracellular Glycoproteins for Crystallization

DOI:
10.3791/53445-v

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07:08 min

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December 23, 2015

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, ,
1Molecular Microbiology and Microbial Pathogenesis Program,Washington University School of Medicine, 2Department of Internal Medicine,Washington University School of Medicine, 3Drug Discovery Program in Pulmonary and Critical Care Medicine,Washington University School of Medicine, 4Biochemistry Program,Washington University School of Medicine, 5Center for the Investigation of Membrane Excitability Diseases,Washington University School of Medicine, 6Department of Biochemistry and Molecular Biophysics,Washington University School of Medicine, 7Department of Cell Biology and Physiology,Washington University School of Medicine

Chapters

  • 00:05Title
  • 01:02Large-scale Culture and Transient Transfection of 293F Cells
  • 02:40Purification
  • 04:58Results: Expression and Purification of Secreted Mammalian Proteins
  • 06:29Conclusion

Summary

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This is a quick, cost-efficient protocol for the production of secreted, glycosylated mammalian proteins and subsequent single-step purification with sufficient yields of homogenous protein for X-ray crystallography and other biophysical studies.

Tags

Keywords: Mammalian Cell ExpressionExtracellular GlycoproteinsCrystallizationTransient TransfectionNickel Affinity ChromatographyProtein ProductionProtein YieldGlycan MaturationX-ray Crystallography

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