Price was born, the son of Marguerite Cobb (née Willcox) and Vincent Leonard Price, Sr., who was the president of the National Candy Company. His grandfather, Vincent Clarence Price, invented "Dr. Price's Baking Powder", the first cream of tartar baking powder, and secured the family's fortune.

Price attended St. Louis Country Day School. He was further educated at Yale in art history and fine art. He was a member of Alpha Sigma Phi Fraternity and the Courtauld Institute, London. He became interested in theater in the 1930s, appearing professionally on stage from 1935.

He made his film debut in 1938 with Service de Luxe and established himself as a competent actor, notably in Laura (1944), opposite Gene Tierney, directed by Otto Preminger. He also played Joseph Smith, Jr. in the movie Brigham Young (1940), as well as a pretentious priest in The Keys of the Kingdom (1944).

Price's first venture into the horror genre was in the 1939 Boris Karloff film Tower of London in which his character was murdered by Karloff's. The following year he portrayed the title character in the film The Invisible Man Returns (a role he reprised in a vocal cameo at the end of the 1948 horror-comedy spoof Abbott and Costello Meet Frankenstein).

In 1946 Price reunited with Gene Tierney in two notable films, Dragonwyck and Leave Her to Heaven. There were also many villainous roles in slick film noir thrillers like The Web (1947), The Long Night (1947), Rogues' Regiment (1948) and The Bribe (1949) with Robert Taylor, Ava Gardner and Charles Laughton. He was also active in radio, portraying the Robin Hood-inspired crime-fighter Simon Templar, aka. The Saint, in a series that ran from 1943 to 1951.

In the 1950s, he moved into horror films, with a role in House of Wax (1953), the first 3-D film to land in the year's top ten at the North American box office, and then the monster movie The Fly (1958). Price also starred in the original House on Haunted Hill (1959) as the eccentric millionaire Fredrick Loren. (Geoffrey Rush, playing the same character in the 1999 remake, was not only made to resemble Price, but was also renamed Steven Price.) In between these horror films, Price played Baka in The Ten Commandments.

In the 1960s, Price had a number of low-budget successes with Roger Corman and American International Pictures (AIP) including the Edgar Allan Poe adaptations House of Usher (1960), The Pit and the Pendulum (1961), Tales of Terror (1962), The Comedy of Terrors (1963) The Raven (1963), The Masque of the Red Death, and The Tomb of Ligeia (1965).

He also starred in The Last Man on Earth (1964), a film based on the Richard Matheson novel I Am Legend. In 1968 Price gave an iconic, coldly menacing, performance as Matthew Hopkins the "Witchfinder General" in the film of the same name.

He also starred in comedy films, notably the cult-classic Dr. Goldfoot and the Bikini Machine (1965). In 1968 he played the part of an eccentric artist in the musical Darling of the Day opposite Patricia Routledge, displaying an adequate if untrained singing voice.

He often spoke of his pleasure at playing "Egghead" on the Batman television series. Another of his co-stars, Yvonne Craig (Batgirl), often said Price was her favorite co-star. In an often-repeated anecdote from the set of Batman, Price, after a take was printed, started throwing eggs at series stars Adam West and Burt Ward, and when asked to stop replied, "With a full artillery? Not a chance!", causing an eggfight to erupt on the soundstage. This incident is reenacted in the behind-the-scenes telefilm Return to the Batcave: The Misadventures of Adam and Burt.

It was also in the 1960s that he began his role as a guest on the game show The Hollywood Squares, even becoming a semi-regular in the 1970s, including being one of the guest panelists on the finale in 1980. He was known for usually making fun of Rose Marie's age, and using his famous voice to answer maliciously to questions.