Unit

UNIT 5.11 Immunoaffinity Chromatography

  1. Timothy A. Springer

Published Online: 1 MAY 2001

DOI: 10.1002/0471142301.ns0511s05

Current Protocols in Neuroscience

Current Protocols in Neuroscience

How to Cite

Springer, T. A. 2001. Immunoaffinity Chromatography. Current Protocols in Neuroscience. 5:5.11:5.11.1–5.11.11.

Author Information

  1. Center for Blood Research Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: NOV 1998

Abstract

This technique involves elution of a single protein from an immunoaffinity column after prior elution of nonspecifically adsorbed proteins. Antibodies are coupled to Sepharose and the cell lysate is passed over the column. After washing, the specific antigen is eluted from the column using one of a number of methods: brief exposure to either high or low pH, or using the detergent octyl beta-d-glucoside (easily removable by dialysis). All these methods are described in this unit along with a procedure for covalently linking an antibody to Sepharose using the cyanogen bromide activation method.