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Molecular Biology Reports

, Volume 18, Issue 3, pp 223–230 | Cite as

Affinity purification of histidine-tagged proteins

  • Jacky Schmitt
  • Heike Hess
  • Hendrik G. Stunnenberg
Methodology Paper

Abstract

Expression of recombinant proteins is a standard technique in molecular biology and a wide variety of prokaryotic as well as eukaryotic expression systems are currently in use. A limiting step is often the purification of the expressed recombinant protein, particularly if mammalian expression systems that yield low expression levels are employed. Here, we discuss the advantages and restrictions of tagging recombinant proteins with histidines and purifying them by Ni2+-NTA chromatography.

Key words

tagging histidine affinity purification recombinant protein nickel 

Abbreviations

GST

glutathione S-transferase

NTA

nitrilotriacetic acid

His

histidine

PAGE

polyacrylamide gel electrophoresis

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Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Jacky Schmitt
    • 1
  • Heike Hess
    • 1
  • Hendrik G. Stunnenberg
    • 1
  1. 1.Gene Expression ProgrammeEuropean Molecular Biology LaboratoryHeidelbergGermany

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