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Published on: March 21, 2022
Purification of poly-histidine-tagged proteins
Sinéad T Loughran1, Dermot Walls
1School of Biotechnology and National Centre for Sensor Research, Dublin City University, Dublin, Ireland.
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 immobilised metal affinity chromatography (IMAC). Here, we describe efficient procedures for the isolation of highly purified His-tagged target proteins from an Escherichia coli host using IMAC.
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