Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4275

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

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

A rapid method for capturing an antigen on the antibody-bead matrix is to mix the antigen solution with the beads and rotate or rock the slurry, allowing maximum contact between the antigen and the immobilized antibody. After the binding reaction has been completed, the slurry is passed into a column for collection of the beads, and then the antigen is eluted (see Immunoaffinity Purification: Eluting Antigens from Immunoaffinity Columns). Because of the difficulties of moving the slurry to the column, this approach is a little more cumbersome than the column . . . [Full Text of this Article]


MATERIALS

Reagents

Equipment


METHOD


TROUBLESHOOTING


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