Malignant cells have been shown to utilize more glucose than normal cells in vitro and in vivo. Glut3 is a member of the facilitative glucose transporters family of transmembrane proteins, and its mRNA levels has been found to be elevated in human cancers, indicating that it may play a role in glucose uptake by cancer cells. Localization and extent of expression of Glut3 protein in normal and malignant human tissues is still largely unknown. We studied Glut3 expression in a series of 325 benign and malignant human tissues using standard immunoperoxidase technique. Of the normal tissues tested, Glut3 immunoreactivity was detected only in normal testis and placenta. Twelve of 14 (86%) testicular, 3 of 22 (16%) ovarian, 2 of 8 (25%) gastric, and 11 of 41 (27%) non-small cell lung carcinomas were positive for Glut3. Other carcinomas, including those of the breast and colon, were negative. Only in Glut3-positive testicular carcinomas were most tumor cells Glut3-positive. We conclude that a) Glut3 has a limited expression in normal and malignant human tissues, as determined by immunohistochemical staining, b) Glut3 may play a role in glucose uptake in a subset of carcinomas of the lung stomach and ovary, and, therefore, these tumors may have a distinct clinical behavior, and c) Glut3 may be an attractive target for monoclonal therapy or imaging of testicular germ cell tumors.