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Purification Using Affinity Tag Technology

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Advanced Methods in Structural Biology

Part of the book series: Springer Protocols Handbooks ((SPH))

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Abstract

Affinity tag technology is a prerequisite for high and rapid purification of recombinant proteins in structural studies because of specific interactions of tags. Widely used tags are polyhistidine tags specific to metal-chelating ligands and glutathione S-transferase tag for glutathione-immobilized ligands. Furthermore, tags binding to antibodies, such as FLAG, Fc, and HA, are also popular for protein preparation and, in addition, are utilized for biological and biochemical analyses, e.g., western blotting, immunoprecipitation, immunofluorescence assay, and flow cytometry. Some tags improve the solubility of proteins. In this chapter, we introduce the features of these representative tags and show several practical examples.

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Author information

Authors and Affiliations

  1. Laboratory of Biomolecular Science, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan

    Atsushi Furukawa

  2. Laboratory of Biomolecular Science and Center for Research and Education on Drug Discovery, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan

    Katsumi Maenaka

  3. Center for Research and Education on Drug Discovery, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan

    Takao Nomura

Authors
  1. Atsushi Furukawa
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  2. Katsumi Maenaka
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  3. Takao Nomura
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Corresponding author

Correspondence to Atsushi Furukawa .

Editor information

Editors and Affiliations

  1. Institute of Materials Structure Science, High Energy Accelerator Research Organization (KEK) , Tsukuba, Ibaraki, Japan

    Toshiya Senda

  2. Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Hokkaido, Japan

    Katsumi Maenaka

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Furukawa, A., Maenaka, K., Nomura, T. (2016). Purification Using Affinity Tag Technology. In: Senda, T., Maenaka, K. (eds) Advanced Methods in Structural Biology. Springer Protocols Handbooks. Springer, Tokyo. https://doi.org/10.1007/978-4-431-56030-2_4

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  • DOI: https://doi.org/10.1007/978-4-431-56030-2_4

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  • Publisher Name: Springer, Tokyo

  • Print ISBN: 978-4-431-56028-9

  • Online ISBN: 978-4-431-56030-2

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