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Poly-Histidine-Tagged Protein Purification Using Immobilized Metal Affinity Chromatography (IMAC)

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Protein Chromatography

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2699))

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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). In this chapter, we describe efficient procedures for the isolation of highly purified His-tagged target proteins from an Escherichia coli host using IMAC in a bind-wash-elute strategy that can be performed under both native and denaturing conditions.

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Author information

Authors and Affiliations

  1. Department of Life and Health 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, 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 .

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Editors and Affiliations

  1. Department of Life and Health Sciences, Dundalk Institute of Technology, Dundalk, Louth, Ireland

    Sinéad T Loughran

  2. National Institute for Bioprocessing Research and Training (NIBRT), Blackrock, Dublin, Ireland

    John Joseph Milne

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Loughran, S.T., Bree, R.T., Walls, D. (2023). Poly-Histidine-Tagged Protein Purification Using Immobilized Metal Affinity Chromatography (IMAC). In: Loughran, S.T., Milne, J.J. (eds) Protein Chromatography. Methods in Molecular Biology, vol 2699. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3362-5_11

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  • DOI: https://doi.org/10.1007/978-1-0716-3362-5_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3361-8

  • Online ISBN: 978-1-0716-3362-5

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