Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4284

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Purification of Antibodies on an Antigen Column

Ed Harlow and David Lane

This protocol was adapted from "Handling Antibodies," Chapter 4, in Using Antibodies by Ed Harlow and David Lane. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1999.

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


INTRODUCTION

Immunoaffinity purification of antibodies is used to purify antigen-specific antibodies from a preparation of polyclonal antibodies. Such purification is commonly needed in the production of antipeptide antibodies, where it is used to concentrate the desired antibodies and separate them from those raised against carrier proteins. It is also used for the more general purpose of removing unwanted, nonspecific binding activity from polyclonal antibody preparations. This method is not appropriate for monoclonal antibodies, which are already homogeneous in their antigen-binding activity.


MATERIALS

Reagents

Antibody serum

Antigen

caution Ethanolamine (100 mM, pH 7.5)

caution Glycine (100 mM, pH 2.5)

caution Merthiolate

recipe caution Sodium phosphate (0.05 M, pH 7.5), . . . [Full Text of this Article]

Equipment


METHOD


TROUBLESHOOTING


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