Peanut agglutinin (PNA)-binding sites in human prolactin (PRL)-producing pituitary adenomas were examined by light and electron microscopy together with immunoblot analysis. At the light microscopic level, the majority of the PRL-producing adenoma cells stained positively for PNA in 15 of 20 cases. PNA binding observed in the cytoplasm had a granular appearance. PRL-producing cells adjacent to the adenoma tissue showed negative PNA staining. In normal pituitary glands, the PRL-positive glandular cells were negative for PNA staining. By electron microscopy, reaction products showing PNA-binding sites were detected in some of the secretory granules. Immunoblotting analysis revealed that the PRL bands corresponded to PNA-stained ones with the exception of the main 23-kDa band. PNA-binding sites have some relation to the secretory granules containing glycosylated forms of PRL. These observations suggest that PNA staining can be used as a valuable method to analyze human PRL-producing pituitary adenomas.