Leon Ames was born on January 20, 1902, in Portland, Indiana, son of Charles Elmer Wycoff and his wife, Cora Alice DeMoss. Some sources list his original last name as "Wykoff" or "Waycoff", and in his early films, he acted under the name Leon Waycoff. In 1935, Ames explained that he changed his name because Waycoff was often misspelled and mispronounced. Ames was his mother's maiden name. In the 1910 census, when his family was residing in Fowler, Indiana, Ames' name is given as Harry L. Wycoff. His father Charles was employed as managing a meat market.
Ames' involvement with entertainment began when he worked as stage manager for the Charles K. Champlin Theatre Company. He ventured into acting with the group and progressed to having the lead in a production of Tomorrow and Tomorrow in Los Angeles. He acted for three years with the Stuart Walker Stock Company in Cincinnati, Ohio.
He debuted on Broadway in It Pays to Sin (1933). His other Broadway credits include Howie (1958), Winesburg, Ohio, (1958), Slightly Married (1943), The Russian People (1942), Little Darling (1942), Guest in the House (1942), The Land Is Bright (1941), The Male Animal (1940), Thirsty Soil (1937), A House in the Country (1937), and Bright Honor (1936).

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