The Duquesne Brewing Company was founded in 1899; On July 5 of that year, the city of Pittsburgh granted the company the right to build a railroad siding to its new plant. 
Before Prohibition, Pittsburgh's brewing operations had begun to combine into fewer players, with most ending up under either the Pittsburgh Brewing Company or Independent Brewing Company of Pittsburgh umbrellas. Duquesne was among the latter, one of 15, in 1905.
Prohibition, starting in 1920, forced many breweries, distillers, and taverns to close. Duquesne was one of only 725 American breweries left when the movement was repealed in April 1933. The company, reformed under the idle charter of Duquesne Brewing Company, emerged as the largest brewing company in Pennsylvania at 325,000 barrels. The South Side brewery, Plant Number 1, was issued IRS U-Permit PA-U-319. In 1937, the company renamed itself to the Duquesne Brewing Company of Pittsburgh.
Duquesne's production capacity increased to two million barrels after World War II when a new building opened at the South Side site in 1950, making it one of the top ten breweries in the United States. The company's best known brand was "Duke," and its popular advertising slogan was "Have a Duke!" Per Kern v. Duquesne Brewing Co. of Pittsburgh (152 A.2d 682, 396 Pa. 279, 1959) the company had plants in Carnegie (Plant 3) and McKees Rocks (Plant 2) in addition to the South Side flagship brewery, although those closed after the new brew house opened at Plant 1 in 1950, with Plant 3 closing in 1952, and Plant 2 in 1950. The curved profile of the 1950 building at Plant 1 was to accommodate the PRR Whitehall Branch, which serviced the brewery from sidings along and off Mary Street. In January 1963, Duquesne expanded into the Cleveland market by purchasing the rights to P.O.C. from Pilsener Brewing Co. after they closed their plant.  P.O.C., which formerly was the Pride of Cleveland, became Pleasure on Call.
While the company had been profitable through the 50s, profits had declined by 1962 to $211,586 and the trend showed no sign of abating . A proposed 1965 acquisition by Pittsburgh Brewing ran afoul of anti-trust laws, and was enjoined in United States v. Pittsburgh Brewing Co. and Milton G. Hulme. Longtime stewards the Friday family would lose control to investor Raymond Sigesmund in a stock battle in 1966. Sigesmund brought in his son-in-law, Franklyn D. Jeans, to run the brewery, but the problems continued. Jeans fired Lando, Inc., the company which had replaced long-time agency Vic Maitland in 1965 on the Duquesne Brewing advertising account, and replaced them with his own company, Admark. Duquesne Brewing, due to competition from national brands and labor problems, sold its labels to C.L. Schmidt Brewing Company of Philadelphia in late 1972, and closed the flagship plant. Schmidt brewed Duquesne in Cleveland into the 1980s before giving up the brand. Lawsuits from displaced employees , as well as defamation suits brought by the former management against some lawyers involved in the case, followed.
The Duquesne Brewery's stable of brands included Silver Top Lager, Silver Top Ale, Duke Ale, Duquesne Pilsener, Duquesne Bavarian Beer, Duquesne Bock Beer, Duquesne Heavy-Dry Beer, Duquesne Draft Beer, Duquesne Porter, Old Nut Brown Ale, Frontenac Pale Ale, Duquesne Beer, Carnegie beer and Duquesne Buccaneer Beer.
The Duquesne Brewery clock, a Pittsburgh landmark, was the largest single face clock in the world when it was installed elsewhere in Pittsburgh in 1933, advertising a "nationally known soft drink". The 60-foot-by-60-foot clock face, with a 35 foot minute hand and a 25 foot hour hand, both of laminated aluminum, is nearly twice the size of London's Big Ben, and was built in Georgia by Audichron for $12,500 and shipped to Pittsburgh. The clock, designed by Audichron founder John L. Franklin, is driven by a 1.25 horsepower Janett motor. It most likely became the Duquesne Brewery clock in May, 1961 after being removed from the hillside billboard in that April. Images obtained from USGS EarthExplorer of the neighborhood on August 1, 1959, do not show the clock's octagonal shadow. Pictures of the Mount Washington hillside about 20 blocks west of the brewery show the clock attached to a billboard as late as the end of 1954 advertising other beverages, including Coca-Cola in 1937, Ballantine around 1949, and Schlitz at the end of 1954.
The 1899 Duquesne Brewery brewhouse building, located at Mary Street and 21st Street in the Southside Flats neighborhood, now is home to The Brew House Association, which provides housing, studio space, and a gallery for Pittsburgh artists, although building code issues forced the building to be temporarily vacated in September 2009 . The adjacent 1950 brewhouse building has been partially restored but is currently vacant. The cellar and keg department is now Storexpress self storage. A storage and garage building has become living space. The former office building has become Beitler-McKee Optical. The bottling works has become U.S. Cargo. Raff Printing and Rynn's Luggage occupy the case storage and shipping buildings. The bridge between the bottling works and case storage stands, while the bridge connecting the two brewhouses has been dismantled. The Pittsburgh Brewing Co. paid $44,000 to repair the clock when it took over in 1999, paying $5,000 a month to show its logo on the face. In 2002, Equitable Gas paid to have their name placed on the clock. In October 2009, AT&T took over the rights to advertise on the clock and has redesigned the face to display the traditional blue and white AT&T logo.