Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4261

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Optimization of Imidazole Concentrations for Immobilized Metal-Ion Affinity Chromatography

Lars Haneskog

This protocol was adapted from "Immobilized Metal-Ion Affinity Chromatography," contributed by Lars Haneskog, Chapter 12, in Purifying Proteins for Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA, 2004.

The first 15% of the full text of this article appears below.


INTRODUCTION

Most samples from which histidine-tagged proteins are to be purified also contain endogenous protein contaminants that bind to the immobilized metal-ion affinity chromatography (IMAC) adsorbent. Usually, these proteins bind more weakly than the histidine-tagged protein. Optimization of the imidazole concentrations within the sample, binding buffer, and elution buffer can reduce or eliminate the coadsorption or coelution of contaminants . . . [Full Text of this Article]


MATERIALS

Reagents

Equipment


METHOD


TROUBLESHOOTING


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