A monoclonal antibody recognizing a novel murine macrophage glycoprotein antigen (MAG) was prepared by the hybridoma technique. Using live and permeabilized macrophages to estimate surface and total MAG, respectively, it was found that at least 50% of the total antigen was intracellular. In contrast, another macrophage surface glycoprotein antigen, Mac-1, was undetectable in the intracellular pool. Immunofluorescence studies confirmed the existence of a substantial intracellular pool of MAG antigen. Similar results were obtained with a panel of cultured tumour cell lines. In one such cell line, it was shown that surface MAG existed in a distinct punctate pattern indicative of microdomains, whereas surface Mac-1 antigen gave a uniform distribution. The possible role of the surface microdomains in the differential expression of the two surface glycoproteins in intracellular membranes is discussed.

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