Unit

UNIT 5.10 Metal-Chelate Affinity Chromatography

  1. Kevin J. Petty

Published Online: 1 MAY 2001

DOI: 10.1002/0471142301.ns0510s05

Current Protocols in Neuroscience

Current Protocols in Neuroscience

How to Cite

Petty, K. J. 2001. Metal-Chelate Affinity Chromatography. Current Protocols in Neuroscience. 5:5.10:5.10.1–5.10.15.

Author Information

  1. Merck & Company, West Point, Pennsylvania

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: NOV 1998

Abstract

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 (Ni2+) 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.