From 1993 until 2004, she played Roz Doyle on the sitcom Frasier, a spin-off of Cheers, starring Kelsey Grammer as Frasier Crane, the role he had played on Cheers since 1984. Along with the principal cast, she won a Screen Actors Guild Award for Outstanding Performance by an Ensemble in a Comedy Series in 1999.
In 2009, she appeared in the ABC Family drama Make It or Break It, for which she received a Gracie Award in the category of Outstanding Female Actor in a Supporting Role in a Drama Series.
Gilpin and Frasier co-star Jane Leeves ran a production company, Bristol Cities (named from the cockney rhyming slang), whose projects included a Fox network pilot titled Minister of Divine, an American remake of the British sitcom The Vicar of Dibley, with Kirstie Alley (another Cheers regular) in a starring role.
She is the voice artist for the character Jane Proudfoot in Final Fantasy: The Spirits Within. She also provided the voices of Desiree in the Nickelodeon animated television series Danny Phantom, Volcana in Superman: The Animated Series and Justice League, Hecate in Hercules: The Animated Series, and Kate Corrigan in the Hellboy animated films. She also did voice work for several Wells Fargo and Johnsonville Meats TV commercials.
She had a recurring role in Men at Work on TBS as editor of Full Steam magazine. In the second season of Scorpion, she was introduced as Katherine Cooper, a team superior in homeland security.