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A High Yield and Cost-efficient Expression System of Human Granzymes in Mammalian Cells
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Biologia
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JoVE Journal Biologia
A High Yield and Cost-efficient Expression System of Human Granzymes in Mammalian Cells

A High Yield and Cost-efficient Expression System of Human Granzymes in Mammalian Cells

DOI:
10.3791/52911-v

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09:16 min

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June 10, 2015

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, , , , ,
1Cellular and Molecular Medicine Program,Boston Children’s Hospital and Harvard Medical School, 2Department of Medicine,University of Fribourg, 3Centre Médical Universitaire,University of Geneva

Capítulos

  • 00:05Título
  • 01:40Expression of Gzms in HEK293T Cells
  • 03:52Purification of Gzms from Culture Supernatant by Nickel-IMAC
  • 05:25Enterokinase (EK) Treatment and Final Clean-up by Cation Exchange Chromatography
  • 07:46Results: Analysis of Recombinant Granzymes by SDS-PAGE and Colorimetric Assays
  • 08:47Conclusion

Summary

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Tadução automática

We describe here a cost-efficient granzyme expression system using HEK293T cells that produces high yields of pure, fully glycosylated and enzymatically active protease.

Tags

Keywords: GranzymesCytotoxic T LymphocytesNatural Killer CellsPerforinGranulysinProtein ExpressionMammalian CellsHEK293TAffinity ChromatographyEnterokinaseCation Exchange ChromatographyGzm AGzm BGzm M

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