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Visualization of anti-sperm plasma membrane IgG and Fab as a method for localization of boar sperm membrane antigens

LD Russell, RN Peterson and TA Russell

A simple method for ultrastructural localization of sperm surface antigens by direct visualization of bound antibodies is presented. Anti- sperm plasma membrane (ASPM) immunoglobulin (Ig) G, visualized in tissues treated with an osmium:ferrocyanide mixture, projected 11-13 nm from the surface and ASPM Fab fragments projected 8-10 nm from the surface. The density of IgG labeling, as subjectively estimated, corresponded to indirect immune fluorescein isothiocyanate, indirect immunoferritin, and sperm-vesicle labeling patterns. Agglutination of sperm vesicles and sperm were demonstrated and the linking antibody visualized. A second antibody on protein A directed against ASPM IgG made the immunologic tag more apparent and indicated, in disrupted sperm preparations, labeling of both sides of the plasma membrane. The method provides for easy and sensitive localization of sperm surface antigens at the ultrastructural level and is presently being used to localize specific sperm antigens.

Volume 30, Issue 12, pp. 1217-1227, 12/01/1982
Copyright © 1982 by The Histochemical Society


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