Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4262

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Protein Purification Using Immobilized Metal-Ion Affinity Chromatography under Optimized Imidazole Concentrations

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

Perform this protocol after determining the imidazole concentration that yields the best purification of the recombinant protein of interest (see Optimization of Imidazole Concentrations for Immobilized Metal-Ion Affinity Chromatography).


MATERIALS

Reagents

Binding, wash, sample and elution buffers (see Step 1)

recipe IMAC high imidazole buffer

recipe IMAC buffer P containing 20 mM . . . [Full Text of this Article]

Equipment


METHOD


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