This protocol describes protein production in mammalian cells by transient transfection. It assumes the expression construct contains either a 6-HIS or Fc fusion tag to allow recovery of the protein by affinity chromatography. The method is one of the simplest available for protein expression in eukaryotic cells, requires little specialized equipment, and has a reasonably high rate of success.

Original languageEnglish
Pages (from-to)107-116
Number of pages10
JournalMethods in Molecular Biology
StatePublished - 2014


  • CMV expression vector
  • HEK293-F cells
  • Kifunensine
  • Kozak
  • Mammalian cell
  • PEI
  • Polyethylenimine
  • Signal sequence
  • Transient transfection


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