We investigated the distribution of protein disulfide isomerase (PDI) family proteins PDI, ERp72, and ERp61 in rat tissues and compared their localization by immunohistochemical double staining. The extent of their expression was diverse in different cell types, although they were ubiquitously distributed in a wide variety of cell types. Prominent staining for all three proteins was detected in thyroid follicular epithelia, tracheal mucous gland cells and chondrocytes, and chief cells and mucous neck cells of the stomach. Hepatocytes, neuronal cells in brain and spinal cord, and pancreatic acinar cells were also consistently and uniformly stained, but with low intensity. On the other hand, distinct differences were observed for the expression of the three proteins in plasma cells, pancreatic islet cells, goblet cells, and Paneth's cells of intestines, seminiferous epithelia, and salivary gland ductal epithelia. The similarities and differences in the distribution of the three proteins provide helpful clues to their biological functions.