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Marlowe was born Hugh Herbert Hipple in Philadelphia, Pennsylvania, and began his stage career in the 1930s at the Pasadena Playhouse in California. Marlowe was usually a secondary lead or supporting actor in the films he appeared in.

His films included Meet Me in St. Louis (1944), Twelve O'Clock High (1949), All About Eve (1950), The Day the Earth Stood Still (1951), Howard Hawks' Monkey Business (1952), Earth vs. the Flying Saucers (1956), Elmer Gantry (1960), Birdman of Alcatraz (1962) and Seven Days in May (1964).

Marlowe was also a regular on the daytime television soap opera, Another World, the last of four actors to portray Matthews family patriarch Jim Matthews, from 1969 until his death from a heart attack, at age 71, in 1982.