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Immunological Investigations
A Journal of Molecular and Cellular Immunology
Volume 22, 1993 - Issue 1
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Original Article

Cell Membrane Antigen-Antibody Complex Dissociation by the Widely Used Glycine-Hcl Method: An Unreliable Procedure for Studying Antibody Internalization

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Pages 1-12 | Published online: 07 Jul 2009
 

Abstract

Methods following the process of binding and internalization of antibodies to cell surface antigens have often employed low pH isoosmolar buffers in order to dissociate surface antigen-antibody complexes. One of the most widely used buffers is a 0.05 M glycine-HCL buffer pH 2.8. Since the efficacy of action of this buffer was critical to a series of internalization experiments employing monoclonal antibodies (Mabs) to carcinoembryonic antigen (CEA) expressing cancer cell lines in this laboratory, we tested its performance in a number of different assays. Our results indicate that this buffer only partially dissociates antigen-antibody bonds and therefore can introduce major inaccuracies in internalization experiments.

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