title: 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 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.
authors.affiliation: Department of Applied Sciences, School of Health and Science, Dundalk Institute of Technology, Dundalk, Louth, Ireland. sinead.loughran@dkit.ie.
authors.forename: Sinéad T
authors.initials: ST
authors.lastname: Loughran
authors.affiliation: Department of Applied Sciences, School of Health and Science, Dundalk Institute of Technology, Dundalk, Louth, Ireland.
authors.forename: Ronan T
authors.initials: RT
authors.lastname: Bree
authors.affiliation: School of Biotechnology, Dublin City University, Dublin 9, Ireland.|National Centre for Sensor Research, Dublin City University, Dublin 9, Ireland.
authors.forename: Dermot
authors.initials: D
authors.lastname: Walls
mesh_terms: D015153:Blotting, Western; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006639:Histidine / Q000737:chemistry / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D012995:Solubility
chemical_list: D011993:Recombinant Fusion Proteins; C033223:polyhistidine; D006639:Histidine
keywords: 6×Histidine; Affinity chromatography; IMAC; Ni-NTA; Polyhistidine; Protein purification