title: Affinity purification of histidine-tagged proteins.
abstract: Expression of recombinant proteins is a standard technique in molecular biology and a wide variety of prokaryotic as well as eukaryotic expression systems are currently in use. A limiting step is often the purification of the expressed recombinant protein, particularly if mammalian expression systems that yield low expression levels are employed. Here, we discuss the advantages and restrictions of tagging recombinant proteins with histidines and purifying them by Ni(2+)-NTA chromatography.
authors.affiliation: European Molecular Biology Laboratory, Gene Expression Programme, Heidelberg, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Schmitt
authors.forename: H
authors.initials: H
authors.lastname: Hess
authors.forename: H G
authors.initials: HG
authors.lastname: Stunnenberg
mesh_terms: D000818:Animals; D001665:Binding Sites; D002846:Chromatography, Affinity / Q000379:methods; D005069:Evaluation Studies as Topic; D006367:HeLa Cells; D006639:Histidine / Q000737:chemistry; D006801:Humans; D009532:Nickel; D009571:Nitrilotriacetic Acid; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D011994:Recombinant Proteins; D006639:Histidine; D009532:Nickel; D009571:Nitrilotriacetic Acid