Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4274

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Immunoaffinity Purification: Binding of Antigen to Antibody-Bead Matrix in a Column

Ed Harlow and David Lane

This protocol was adapted from "Immunoaffinity Purification," Chapter 9, in Using Antibodies by Ed Harlow and David Lane. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1999.

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


INTRODUCTION

The simplest method to bind the antigen to the antibody-bead matrix is to collect the beads in a column and pass the antigen solution down the column. The efficiency of this method depends on the contact time between the immobilized antibody and the antigen, which can be extended by using low flow rates. The immobilized antibody captures the antigen as it flows past and holds it until this interaction is broken in the . . . [Full Text of this Article]


MATERIALS

Reagents

Equipment


METHOD


TROUBLESHOOTING


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