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Purification of Polyhistidine-Tagged Proteins

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Protein Chromatography

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1485))

Abstract

His-tagging is the most widespread and versatile strategy used to purify recombinant proteins for biochemical and structural studies. Recombinant DNA methods are first used to engineer the addition of a short tract of poly-histidine tag (His-tag) to the N-terminus or C-terminus of a target protein. The His-tag is then exploited to enable purification of the “tagged” protein by Immobilized Metal Affinity Chromatography (IMAC). Here, we describe efficient procedures for the isolation of highly purified His-tagged target proteins from an E. coli host using IMAC.

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Author information

Authors and Affiliations

  1. Department of Applied Sciences, School of Health and Science, Dundalk Institute of Technology, Dundalk, Louth, Ireland

    Sinéad T. Loughran & Ronan T. Bree

  2. School of Biotechnology, Dublin City University, Dublin 9, Ireland

    Dermot Walls

  3. National Centre for Sensor Research, Dublin City University, Dublin 9, Ireland

    Dermot Walls

Authors
  1. Sinéad T. Loughran
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  2. Ronan T. Bree
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  3. Dermot Walls
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Corresponding author

Correspondence to Sinéad T. Loughran .

Editor information

Editors and Affiliations

  1. School of Biotechnology, Dublin City University, Dublin, Ireland

    Dermot Walls

  2. Department of Applied Sciences, Dundalk Institute of Technology, Dublin, Ireland

    Sinéad T. Loughran

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Loughran, S.T., Bree, R.T., Walls, D. (2017). Purification of Polyhistidine-Tagged Proteins. In: Walls, D., Loughran, S. (eds) Protein Chromatography. Methods in Molecular Biology, vol 1485. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6412-3_14

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  • DOI: https://doi.org/10.1007/978-1-4939-6412-3_14

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6410-9

  • Online ISBN: 978-1-4939-6412-3

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