The localization of the alpha subunit of the S-100 protein (S-100 alpha) and beta subunit (S-100 beta) was studied in the peripheral nervous system of the rat. In peripheral nerves, S-100 alpha and S-100 beta were found in the cytoplasm of Schwann cells. Axons were positively stained in part by S-100 alpha and almost totally by S-100 beta. In the dorsal root ganglia, S-100 alpha was found in satellite cells and their processes and in some neurons. S-100 beta was found in more of the large neurons, but almost all of the small neurons were negative for S-100 beta. In the anterior horn cells, S-100 beta staining was stronger than that of S-100 alpha. In Schwann cells, both S-100 alpha and S-100 beta were present on the rough endoplasmic reticulum, free ribosomes, and nucleus, as seen by electron microscopy. The S-100 alpha and S-100 beta in axons were associated with microtubules and neurofilaments.