Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4197

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Packing a Size-Exclusion Column

Helena Hedlund

This protocol was adapted from "Size-Exclusion Chromatography," contributed by Helena Hedlund, Chapter 7, in Purifying Proteins for Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2004.

The first 100 words of the full text of this article appear below.


INTRODUCTION

Size-exclusion columns can be packed in various ways depending on the type of column and equipment used. Always read and follow the relevant column instruction manual carefully. How well the column is packed will have a major effect on the result of the separation. The following method can be used for packing Superdex Prep grade media into an XK 16/60 (16 x 600 mm) column (Amersham Biosciences).


MATERIALS

Reagents

caution Acetone or 0.2 M NaCl (see Step 13)

Equipment

Chromatography column, XK 16/60 column

Chromatography pump

Glass-fiber filter

Packing reservoir, RK 16

Superdex Prep grade media

UV or conductivity monitor


METHOD

Preparation of the Gel . . . [Full Text of this Article]


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