We have utilized monoclonal antibodies to investigate the antigenic diversity of MAP2-immunoreactive proteins in the nervous system of vertebrates. We found that domains defined by the monoclonal antibodies differed in their conservation across vertebrate evolution, ranging from wide cross-reactivity with almost all vertebrates (mammals, birds, reptiles and amphibians) to a very limited cross-reactivity with only few mammalian species. However, we did not find MAP2-immunoreactive proteins in fish species with either of the monoclonal or polyclonal antibodies. There was also a significant divergence in the apparent molecular weight of MAP2, even in closely related species. For example, different species of wild mice and strains of laboratory mice showed variations of up to 30 kDa in their apparent molecular mass. Using alkaline phosphatase, under conditions that dephosphorylate neurofilaments, we showed that the observed heterogeneity was not the result of variations in the phosphate content. The heterogeneity in molecular weight of MAP2 may, therefore, be the result of changes in primary structure, transcriptional variations or different post-translational modifications. The heterogeneity of MAP2, as well as its specific distribution and implicated interactions with other molecules, underscore the complexity of MAP2 and its potential for structural and functional diversity. The phylogenic analysis of such a complex molecule also provides a method to establish the uniqueness of monoclonal antibodies and the degree of their conservation for their corresponding epitopes.