Detection of guinea pig macrophages by a new CD68 monoclonal antibody, PM-1K
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A new monoclonal antibody, PM-1K, was raised against 24-h cultured human peritoneal macrophages. In immunohistochemical assays, PM-1K recognized freshly isolated blood monocytes and most tissue macrophages as well as myeloid dendritic cells such as Langerhans cells and interdigitating cells. The molecular size of the antigen recognized by PM-1K was determined to be 110 kD by means of immunoaffinity purification. Because this affinity-purified antigen recognized by PM-1K was also recognized by anti-CD68 antibodies, it is believed to be one of the heterogeneous molecules of the CD68 antigen. Analysis showed interspecies reactivity of PM-1K with macrophages from guinea pigs, pigs, bovine species, and monkeys. Among these macrophages, those of the guinea pig reacted strongly with PM-1K. Patterns of PM-1K immunostaining in guinea pig tissues were similar to those found in human tissues. Studies with the immunoelectron microscope revealed reaction products of PM-1K in the cytoplasm, especially around endosomes. Since only a few antibodies are available to label guinea pig macrophages, PM-1K is considered to be one of the most suitable antibodies to examine macrophages in experimental guinea pig models.
KeywordsCD68 Macrophages Guinea pigs Immunohistochemistry Monoclonal antibody
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We thank Dr. Araki Norie (Department of Tumor Genetics and Biology, Kumamoto University, Kumamoto, Japan) for helpful discussions during the course of this work; Dr. Hisao Saitoh (Department of Regeneration Medicine, Institute of Molecular Embryology Genetics, Kumamoto University, Kumamoto, Japan) for technical advice; and Takenobu Nakagawa, Osamu Nakamura, Makiko Komohara, and Jun-ichi Yoshida for technical assistance.
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