Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4267

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Packing Ceramic Hydroxyapatite Columns for High-Performance Liquid Chromatography

Richard J. Simpson

This protocol was adapted from "Protein Chromatography on Hydroxyapatite," contributed by Richard J. Simpson, Chapter 13, 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

Ceramic hydroxyapatite is a spherical, macroporous form of hydroxyapatite that overcomes many of the limitations of crystalline hydroxyapatite (especially mechanical instability), that, as a result, can be used reproducibly at high flow rates and high pressures in large columns. This protocol, adapted with permission from the Bio-Rad application notes for CHT ceramic hydroxyapatite, describes the packing of ceramic hydroxyapatite for HPLC.


MATERIALS

IMPORTANT:

Use carbonate-free H2O to prepare all solutions. Carbonate reacts with calcium ions to form a crust of calcium carbonate at the top of the column.

Reagents

CHT ceramic hydroxyapatite (Bio-Rad)

Because of its ceramic nature, ceramic hydroxyapatite is . . . [Full Text of this Article]

Equipment


METHOD


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