To identify somite-specific antigens exhibiting novel expression patterns in the developing chick embryo, epithelial somites were isolated and used for intrasplenic immunization. Immunohistochemical screening of chick embryos of various stages focussed our attention on a monoclonal antibody, Som5H5, which stained somitic derivatives, spinal nerves, and neural tubes as well as the costameres of adult skeletal muscle. Western blot and mass spectrometric analysis revealed the Som5H5 antigen to be vimentin, the distribution of which has been well characterized before. In addition to the described vimentin pattern, Som5H5 stained a region in the proximal portion of the developing limb bud. This novel expression domain was confirmed by in situ hybridization using vimentin riboprobes. Signaling molecules (Shh and BMP-2), known to play a role in limb development, did not influence vimentin expression. Thus, no functional or cellular correlate to this vimentin-positive region could be determined.