Antibodies, A Laboratory Manual, Second Edition, Edited by Edward A. Greenfield



Table of Contents

Expand All | Contract All

Purification of Antibodies: High-pH DEAE Chromatography

(Protocol summary only for purposes of this preview site)

If the antibody solution is adjusted to a basic pH of 88.5, then IgG binds to the DEAE resin. After washing the column, the antibody is eluted by adding a buffer of increasing ionic strength to the column. Prepacked columns of many sizes are available for the isolation of antibodies by DEAE chromatography. Alternatively, DEAE medium can be swelled or prepped according to the manufacturer's instructions and a column can be poured when needed. In either case, it is important to choose a column of sufficient size to bind the IgG in your sample. A column that contains 23 mL of DEAE beads per milliliter of serum (or equivalent) is sufficient for most applications.

Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition

Search for information about other protocols included in the book: