Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4215

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Packing an IMAC Column

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, NY, USA, 2004.

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


INTRODUCTION

This protocol describes a procedure for packing a 30-ml immobilized metal-ion affinity chromatography (IMAC) column with the aid of a pump.


MATERIALS

IMPORTANT:

Equilibrate all materials to room temperature prior to use. Use double-distilled H2O, or similar quality, for all of the packing steps.

Reagents

Ethanol

caution NiSO4 (0.1 M)

Equipment

Column (1.6 cm I.D. x 15 cm), empty with suitable flow adapters, and inlet and outlet ports

Glass filter (100-ml)

IMAC resin (e.g., Chelating Sepharose Fast Flow, Amersham Biosciences)

Packing reservoir

Pump


METHOD

1. Gently . . . [Full Text of this Article]


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