Purification of polyhistidine-tagged proteins

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 Histag 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 Histagged target proteins from an E. coli host using IMAC.