A Monoclonal Antibody Generated against Chick Retina Cell Membrane: Cell Selectivity and Properties of the Antigen
Andreae, Margie Clare
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A monoclonal antibody has been produced against a membrane preparation from chick embryonic retina. This hybridoma antibody, termed 5A-II, was characterized for its cell selectivity and developmental distribution pattern in the avian retina using immunohistochemical methods on intact and dissociated tissue. Results showed a uniform staining pattern on pre-differentiated tissue becoming specific preferentially to Muller glia cells, and somewhat to photoreceptor cell surface in the mature neural retina. Initial tests were done to determine the biochemical nature of the antigen. Cell membrane preparations were exposed to various proteases, detergents and other identifying reagents, and then immunostained with 5A-II to test for any effect on immunoreactivity. The antigen was insensitive to protease treatment and, at the same time, highly sensitive to the detergents, suggesting the antigen is lipid in nature. In conclusion, monoclonal antibody 5A-II is preferentially specific to Muller cell surfaces in mature avian retina, and its antigenic determinant is most likely a membrane lipid.