Metal-chelate affinity chromatography.
Recombinant proteins engineered to have six consecutive histidine residues on either the amino or carboxy terminus can be purified using a resin containing nickel ions (Ni(2+)) that have been immobilized by covalently attached nitrilotriacetic acid (NTA). This technique is know as metal-chelate affinity chromatography and can be performed using either native or denatured protein. This unit presents protocols for expression of histidine-tail fusion proteins and their purification in either native or denatured form (along with procedures for renaturation by either dialysis or solid-phase renaturation). Also provided are procedures for analysis of the purified produce and regeneration of the NTA resin.
Petty
K J
KJ
Merck & Company, West Point, Pennsylvania, USA.
D000818:Animals; D002413:Cations, Divalent / Q000032:analysis; D002614:Chelating Agents / Q000032:analysis* / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D008670:Metals / Q000032:analysis* / Q000737:chemistry; D009532:Nickel / Q000032:analysis / Q000737:chemistry; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry
D002413:Cations, Divalent; D002614:Chelating Agents; D008670:Metals; D011994:Recombinant Proteins; D009532:Nickel