Protocol

Preparation of Denatured Protein Samples from Inclusion Bodies

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.

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INTRODUCTION

To prepare inclusion bodies from Escherichia coli, cells are lysed, cell debris is separated from inclusion bodies by centrifugation, and the inclusion bodies are washed in a low concentration of denaturant in the presence of mild detergents. The inclusion bodies may be stored in a buffer without denaturants and detergent. Before . . . [Full Text of this Article]

MATERIALS

Reagents

Equipment

METHOD