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Transient Expression in HEK 293 Cells: An Alternative to E. coli for the Production of Secreted and Intracellular Mammalian Proteins

  • Joanne E. NettleshipEmail author
  • Peter J. Watson
  • Nahid Rahman-Huq
  • Louise Fairall
  • Mareike G. Posner
  • Abhishek Upadhyay
  • Yamini Reddivari
  • Jonathan M. G. Chamberlain
  • Simon E. Kolstoe
  • Stefan Bagby
  • John W. R. Schwabe
  • Raymond J. Owens
Part of the Methods in Molecular Biology book series (MIMB, volume 1258)

Abstract

Transient transfection of human embryonic kidney cells (HEK 293) enables the rapid and affordable lab-scale production of recombinant proteins. In this chapter protocols for the expression and purification of both secreted and intracellular proteins using transient expression in HEK 293 cells are described.

Key words

Mammalian HEK Transient transfection Secreted protein Intracellular protein Protein complexes FLAG purification 

Notes

Acknowledgment

The OPPF-UK is funded by the Medical Research Council, UK (grant MR/K018779/1). P.J.W., L.F., and J.W.R.S. are funded by the Wellcome Trust (grants WT085408 and WT100237). J.M.G.C. is funded by a University of Portsmouth IBBS studentship, and S.E.K. by the University of Portsmouth Research Development Fund. M.P., A.U., and S.B. are funded by the BBSRC, UK (grant BB/J008176/1).

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Joanne E. Nettleship
    • 1
    • 2
    Email author
  • Peter J. Watson
    • 3
  • Nahid Rahman-Huq
    • 1
    • 2
  • Louise Fairall
    • 3
  • Mareike G. Posner
    • 4
  • Abhishek Upadhyay
    • 4
  • Yamini Reddivari
    • 1
    • 2
  • Jonathan M. G. Chamberlain
    • 5
  • Simon E. Kolstoe
    • 5
  • Stefan Bagby
    • 4
  • John W. R. Schwabe
    • 3
  • Raymond J. Owens
    • 1
    • 2
  1. 1.OPPF-UK, Research Complex at HarwellR92 Rutherford Appleton LaboratoriesDidcotUK
  2. 2.Division of Structural Biology, Wellcome Trust Centre for Human GeneticsUniversity of OxfordOxfordUK
  3. 3.Department of BiochemistryUniversity of LeicesterLeicesterUK
  4. 4.Department of Biology and BiochemistryUniversity of BathBathUK
  5. 5.Institute of Biomedical and Biomolecular ScienceUniversity of PortsmouthPortsmouthUK

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