The monoclonal antibody 25F9 recognises a protein of 86 kD on the cell surface and within the cytoplasm of mature macrophages. The antibody is associated with fully differentiated tissue macrophages both in normal and in diseased tissues, independently of the presence or absence of inflammation. The antigen is absent on freshly isolated monocytes and other blood cells. After 6 to 7 days culture human monocytes become positive. Some melanoma and carcinoma cell lines are also positive. Furthermore the monoclonal antibody 25F9 cross reacts with a subpopulation of macrophages of rhesus monkey, pig alveolar macrophages and Kupffer cells. The monoclonal antibody 25F9 is very useful for macrophage phenotyping.
Flow cytometry, Frozen sections, Paraffin sections, Western blot
For immunohistology, flow cytometry and Western blotting dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50.