Connell began her career with her husband, Gordon, entertaining in San Francisco night clubs such as The Purple Onion and The Hungry I. Eventually the couple moved to New York City, where Connell made her Off-Broadway debut in the 1955 revival of The Threepenny Opera, a long-running hit at the Theatre de Lys. In the London production of Once Upon a Mattress, Connell starred as Winifred, the role that Carol Burnett originated in New York.
Connell's most prominent success came in 1966 when she was cast as Agnes Gooch in the original Broadway production of Jerry Herman's Mame. She recreated the role in the 1974 screen adaptation after the film's star, Lucille Ball, became dissatisfied with Madeline Kahn, who had been signed originally to play Gooch.
Only four-foot-eleven, Connell was described as a master of the large comic gesture in The Oxford Companion to American Theatre, which described her as "a tiny woman with a giant, squeaking voice".
Jane Connell was nominated for the Tony Award for Best Featured Actress in a Musical for her performance in Me and My Girl (1986). Additional Broadway credits include New Faces of 1956 (1956); Drat! The Cat! (1965); Dear World (1969), once again supporting Angela Lansbury; the short-lived 1983 revival of Mame, in which Lansbury reprised her 1966 lead role; Lend Me a Tenor (1989); Crazy for You (1992); and Moon Over Buffalo (1995), starring Carol Burnett. Her last Broadway appearance was in the role of pianist Jeannette Burmeister in The Full Monty, succeeding Kathleen Freeman, who died during her run in show.
Jane and Gordon Connell enjoyed extensive theatre careers. They appeared together on Broadway in Lysistrata (November 1972), starring Melina Mercouri in the title role. She appeared in New York City Center Encores! production of Call Me Madam (February 1995), and the Weill Recital Hall of Carnegie Hall concert presentation of Noël Coward's Sail Away (November 1999).
Jane Connell's film roles included Ladybug Ladybug (1963), Kotch (1971), Won Ton Ton, the Dog Who Saved Hollywood (1976), House Calls (1978), Rabbit Test (1978), and Dr. Jekyll and Ms. Hyde (1995). Her television appearances included Bewitched in a memorable turn imitating Queen Victoria (as Queen Hepzibah often uttering the phrase, "We are not amused." to comic effect), Green Acres', All in the Family, Love, American Style, M*A*S*H, Maude, Good Times and Law & Order. She was a regular in the cast of the short-lived 1976 situation comedy The Dumplings. From 1991 to 1994, she had the recurring role of social worker Roberta Domedian on the Family Channel sitcom Big Brother Jake.
She married Gordon Connell, an actor and musician, in 1948. They remained married until her death in 2013.
Jane Connell died on September 22, 2013, aged 87, at the Lillian Booth Actors Home of the Actors Fund in Englewood, New Jersey, from undisclosed causes. She is survived by her two daughters, Melissa and Maggie.