Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4283

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Antibody Purification on Protein A or Protein G Columns

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

Purification of antibodies on protein A or protein G columns is rapid and nearly foolproof, and it provides a fold purification that cannot be reached with other conventional chromatography methods. In general, it is the method of choice for commonly used applications. It is not appropriate, however, for purification of antigen-specific fractions from polyclonal antibody preparations. In this case, an antigen affinity column should be used (Purification of Antibodies on an Antigen Column). This protocol is adapted from Ey et al. (1978).


MATERIALS

Reagents

Crude antibody preparation (serum, tissue culture supernatant, or ascites)

caution Glycine (50 mM, pH 3.0)

caution Glycine (100 . . . [Full Text of this Article]

Equipment


METHOD


TROUBLESHOOTING


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