Born in Rochester, Indiana as the son of a saloonkeeper, Chamberlain was raised mostly by his grandmother after his parents divorced. He spent much of his youth in Chicago. After serving in the U.S. Navy from 1943 to 1946, he attended the Art Institute of Chicago (1951–52) and Black Mountain College (1955–56). At Black Mountain, he studied with the poets Charles Olson, Robert Creeley, and Robert Duncan, who were teaching there that semester. The following year, he moved to New York, where for the first time he created sculpture that included scrap-metal auto parts. Over the course of his prolific career, he had studios in New York, New Mexico, Florida, Connecticut, and finally Shelter Island.

Chamberlain is best known for creating sculptures from old automobiles (or parts of) that bring the Abstract Expressionist style of painting into three dimensions. He began by carving and modelling, but turned to working in metal in 1952 and welding 1953. By 1957, while staying with the painter Larry Rivers in Southampton, New York, he began to include scrap metal from cars with his sculpture Shortstop, and from 1959 onward he concentrated on sculpture built entirely of crushed automobile parts welded together. Far more than just another wrinkle on assemblage Shortstop and subsequent works completely reinvented modeling casting, and volume altering Marcel Duchamp’s notion of the readymade and using the car as both medium and tool. In 1962 Donald Judd wrote, “The only reason Chamberlain is not the best American sculptor under forty is the incommensurability of ‘the best’ which makes it arbitrary to say so.”

By the end of the 1960s, Chamberlain had replaced his signature materials initially with galvanized steel, then with mineral-coated Plexiglas, and finally with aluminum foil. In 1966, he began a series of sculptures made of rolled, folded, and tied urethane foam including sofas. Since returning in the mid-1970s to metal as his primary material, Chamberlain has limited himself to specific parts of the automobile (fenders, bumpers, or the chassis, for example). In 1973, two 300-pound metal pieces by Chamberlain were mistaken for junk and carted away as they sat outside a gallery warehouse in Chicago.

In the early 1980s, Chamberlain moved to Sarasota, Florida, where an 18,000-square-foot warehouse studio on Cocoanut Avenue enabled him to work on a much grander scale than he previously had. Many of the subsequent works Chamberlain made in Florida revert to more volumetric, compact configurations, often aligned on a vertical axis. As seen in the so-called Giraffe series (circa 1982–83), for example, linear patterns cavort over multicolored surfaces—the results of sandblasting the metal, removing the paint, and exposing the raw surface beneath. In 1984, Chamberlain created the monumental American Tableau created for display on the Seagram Building’s plaza.

Assembling intricately cut, painted metal parts, Chamberlain made his first mask, A Good Head and a Half (1991), for a benefit auction for Victim Services in 1991, providing aid to victims of sexual assault. He continued to produce masks throughout the 1990s in his studio on Shelter Island, titling many of them with opus numbers.