Following her breakthrough in the comedy-drama film Coyote Ugly (2000),[3] she starred in Cheaper by the Dozen (2003), its sequel Cheaper by the Dozen 2 (2005), The Prestige (2006), Angel Has Fallen (2019), and as CIA agent Annie Walker in the USA Network spy drama series Covert Affairs (2010–2014), for which she was nominated for a Golden Globe Award for Best Actress – Television Series Drama.

She graduated from Toms River High School North in 1994,[8] and earned a bachelor's degree in theater from Honors Tutorial College at Ohio University in 1998.

The film was met with mixed reviews, but the performances of Perabo, Paré and co-star Mischa Barton were widely praised.

[23] Entertainment Weekly's Owen Gleiberman called her "an actress of glittering ferocity" and her performance "a geyser of emotion".

Her other films include The I Inside (2003), Perfect Opposites (2004), George and the Dragon (2004), The Cave (2005), Imagine Me & You (2005), Edison (2005), and The Prestige (2006).

In 2008, Perabo co-starred with Jamie Lee Curtis in the family comedy film Beverly Hills Chihuahua.

"[28][29] The following year, Perabo was cast as CIA field agent Annie Walker, the lead character in Covert Affairs.

[34] In September 2012, Perabo had a supporting role, alongside Bruce Willis, Joseph Gordon-Levitt, and Emily Blunt, in the time travel thriller film Looper.

In 2020, she had a recurring role as Linda Craft in the Showtime supernatural horror series Penny Dreadful: City of Angels.

In 2021, Perabo joined the cast of Yellowstone as Summer Higgins, an animal rights activist and ally of John (season 4).

[43] She speaks fluent French,[21] advocates for LGBT rights and women's political leadership, and serves on the advisory board of VoteRunLead.

[50][51] Perabo is an advisory board member for The Hometown Project, a non-profit organisation that works to help elect candidates in local races.