Cell Surface Molecules Immunoprecipitation Of

Frank AW Verreck and Frits Koning, Department of Immunohepatology and bloodbank, University Hospital Leiden, Leiden, The Netherlands

Copyright © 1998 Elsevier Ltd. All Rights Reserved.

Immunoprecipitation of cell surface molecules is a quick and reliable method that allows the analysis of cell surface expressed molecules recognized by monoclonal antibodies and by antisera. In addition, using proper solubilization techniques, interactions between cell surface expressed molecules can be defined that give insight into subunit interactions in multichain receptor complexes. Usually lysates of cell surface iodinated cells are used. The principle of immunoprecipitation is based on 1) solubilization of membrane proteins by the use of nonionic detergents, 2) subsequent interaction of specific antibody with the solubilized membrane antigen, and 3) recovery of the antibody-antigen complexes by binding to an insoluble support, thereby allowing washing procedures to remove unbound molecules. Analysis of such immunoprecipitates usually takes place by SDS-PAGE or isoelectric focusing (IEF).

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