Edward Stabler came to Alexandria after learning the apothecary trade with his brother in Leesburg, Virginia. A Quaker and an able merchant, he opened his own business in rented space near South Fairfax and Prince Streets in 1792. An apothecary then functioned as pharmacist, retailer, and adviser: customers could describe symptoms directly, purchase prepared medicines, and buy many household or agricultural goods without a physician's prescription.

Stabler's business prospered. In 1805 he bought the lot at 107 South Fairfax Street and erected the three-story brick building that remains part of the museum. By 1829 he had acquired neighboring 105 South Fairfax and incorporated it into the operation. The shop sold medicines, surgical and dental equipment, soap, perfume, paints, varnish, window glass, garden supplies, artists' materials, combs, brushes, and other products required by a growing port town.

After Edward Stabler died in 1831, his son William continued the family enterprise and involved several brothers and his brother-in-law, John Leadbeater. When William died without children in 1852, Leadbeater purchased the firm. His son Edward later expanded its wholesale and mail-order reach. At its height, the company employed traveling salesmen and supplied hundreds of pharmacies around Washington and across parts of Virginia, West Virginia, and North Carolina.

New regulation, changing medical practice, chain-store competition, and financial strain weakened the firm in the twentieth century. It entered bankruptcy protection in 1916, reorganized, and finally closed in 1933 during the Great Depression. Alexandria citizens and the American Pharmaceutical Association acted quickly to save the buildings, archives, fixtures, bottles, and materials. The Landmarks Society acquired the property, and the restored museum opened in 1939.

The City of Alexandria received the museum and collection in 2006. Today the unusually intact site preserves hand-blown glass, botanicals, equipment, prescription books, ledgers, invoices, and family papers from a business that operated from 1792 to 1933. Famous names such as Martha Washington appear in the records, but the deeper value is the documentation of everyday health, commerce, labor, and changing medical knowledge across more than a century. The collection is also valuable because products remained in place when the business failed. Labels, containers, ingredients, and correspondence can be compared rather than studied as isolated antiques. They reveal changing trade networks, advertising, professional standards, and ideas about illness. They also document the people who filled orders, kept accounts, cleaned rooms, delivered goods, and maintained the family enterprise. This combination of building, objects, and archives gives historians an unusually detailed view of an American pharmacy across several generations.