The Journal of Experimental Medicine
Avanti Polar Lipids
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The Journal of Experimental Medicine, Vol 140, 523-537, Copyright © 1974 by The Rockefeller University Press

ARTICLE

HAPTEN-SANDWICH LABELING : I. A GENERAL PROCEDURE FOR SIMULTANEOUS LABELING OF MULTIPLE CELL SURFACE ANTIGENS FOR FLUORESCENCE AND ELECTRON MICROSCOPY



L. Wofsy 1, P. C. Baker 1, K. Thompson 1, J. Goodman 1, J. Kimura 1, and C. Henry 1

1 From the Department of Bacteriology and Immunology, University of California, Berkeley, California 94720

A hapten-sandwich procedure has been developed for specific labeling of cell surface antigens for fluorescence or electron microscopy. Haptens are azo-coupled to immunoglobulins specific for a cell surface antigen; the hapten-modified cell-bound antibodies can then be visualized by adding fluorescent antihapten antibody, or by adding antihapten antibody followed by hapten-modified markers for electron microscopy. Virus or high molecular weight protein markers are lightly cross-linked before conjugation with hapten to prevent their disruption. Such stable hapten-modified markers, and the accessibility of many different purified anti-azophenyl-hapten antibodies, make it feasible to distinguish more than one membrane antigen in a given labeling experiment. When mouse lymphoid cell populations are labeled with separate markers for Ig and for thymus-associated antigens, many cells exhibit the Ig marker exclusively or the thymic marker predominantly, and some cells are completely free of label.

Submitted on April 2, 1974


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