The Harrison Brothers Hardware store in Huntsville, Madison County, is Alabama's oldest operating hardware store. It was originally established as a tobacco shop by brothers James B. and Daniel T. Harrison in 1879 and was damaged by a fire in 1901. The brothers reopened it as a hardware store in 1902 and retained its turn-of-the-century motif. It remained in operation until 1983, after the last remaining Harrison brother died. The Historic Huntsville Foundation, established in 1974, then purchased the building and operates it as a museum and store run by a staff of volunteers.

In 1983, after the last surviving Harrison brother, John Harrison, died, the business and building were purchased by the historic preservation Historic Huntsville Foundation (HHF), which currently owns and operates it as a historic site and gift shop. Before its reopening, the HHF hosted a fundraising benefit with more than 600 individuals in attendance that helped to finance the store's reopening. All profits made from in-store sales go towards the foundation's preservation efforts as well as activities such as summertime Movies in the Park in Big Spring Park.

Harrison Brothers Hardware is run by HHF volunteers who serve as greeters, sales people, special events workers, and store docents. The west wing of the building serves as an art gallery for local, regional, and national artists and hosts monthly art shows. The second floor of the building includes an area called "The Loft." It provides a meeting space for non-profit and community groups, a multifunctional work space for entrepreneurs and small businesses, and the main office of the Historic Huntsville Foundation. The basement has been used to store and sell a variety of architectural trim and accessories recovered from old buildings.
Harrison Brothers Hardware is located as 124 Southside Square. It is open Monday through Saturday from 9:30 a.m. to 5:00 p.m. A historic marker is located outside. Nearby are Alabama Constitution Village, the EarlyWorks Children's Museum, the Weeden House Museum, and Big Spring Park.