Medaille College's main campus is located in Buffalo, New York, on an attractive urban campus at the intersection of Route 198 (Scajaquada Expressway) and Parkside Avenue. Adjacent to Delaware Park and the Buffalo Zoological Gardens, the campus is easily accessible by car or bus. In addition, Medaille is in close proximity to the Albright-Knox Art Gallery, the Buffalo and Erie County Historical Building, the Buffalo Museum of Science, and the Buffalo and Erie County Public Libraries. The campus is served by the Humboldt-Hospital rapid transit station.

The Amherst branch campus is located in the Centerpointe Corporate Park at 400 Essjay Road. Situated on Essjay, which runs between Sheridan Drive and Evans Street, the campus is easily accessible by Main Street (Route 5) or Sheridan Drive (Route 324).