Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot3340

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Removal of Cross-reactive Antibodies from Antiserum: Affinity Chromatography

Joseph Sambrook and David W. Russell

This protocol was adapted from Molecular Cloning, 3rd edition, by Joseph Sambrook and David W. Russell. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2001

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


INTRODUCTION

This protocol describes a method for removing antibodies that react with bacterially encoded proteins by passing a crude preparation of immunoglobulins through a column containing immobilized bacterial proteins.


MATERIALS

E. coli strain used as host for preparation of expression library

Antibody preparation that is to be used for screening

This protocol works best when using an IgG fraction, prepared by chromatography of the antiserum on protein A-Sepharose.

recipe Cell lysis buffer . . . [Full Text of this Article]


METHOD


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