Purification of recombinant 2XMBP tagged human proteins from human cells

Methods Mol Biol. 2014:1176:209-17. doi: 10.1007/978-1-4939-0992-6_17.


The ability to purify an intact, functional protein or protein complex is an essential step in biochemical characterization studies. Challenges in purification arise when proteins are of low abundance or are unstable resulting in low yields or poor in vitro activity. In this protocol we describe a method to purify active, recombinant human proteins fused to a tandem MBP tag after expression in human 293T cells.

MeSH terms

  • Cell Line
  • Chromatography, Affinity
  • Cloning, Molecular
  • Gene Expression
  • HEK293 Cells
  • Humans
  • Proteolysis
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification*
  • Transfection


  • Recombinant Fusion Proteins