De Pablo co-hosted episodes of the Latin-American talk show Control with former Entertainment Tonight host Carlos Ponce at the age of 15 before attending Carnegie Mellon University to study music and theater.

In 2005, De Pablo was about to make her Broadway debut in The Mambo Kings[10] as Dolores Fuentes, but the show closed after a short trial run in San Francisco.

She had submitted a videotape audition while waiting for The Mambo Kings to move forward and was asked to fly to Los Angeles for a screen test two days after it was canceled.

[4][11] Network executives arranged for her to act alongside series regular Michael Weatherly to test for chemistry during her second audition; he went off script by brushing her hair back and commenting, "You remind me of Salma Hayek".

[14] On July 10, 2013, it was reported by CBS that de Pablo would be leaving NCIS for undisclosed reasons, although she remained long enough to conclude her character Ziva David's storyline at the beginning of season 11.

[15] Later, in a 2016 Q&A session, de Pablo stated that part of the reason why she left the show was because of the direction of her character's arc and feeling that the scripts were not good enough to make her stay.

De Pablo made a surprise unannounced return to the show in the last scene of the season 16 finale, which aired on May 21, 2019, in which Ziva arrives at Gibbs' basement to warn him that his life is in danger.

[20] In March 2016, Deadline Hollywood reported de Pablo is slated to return to series television as Laura Kale in Syfy's thriller-drama, Prototype, pending negotiations.

[22] The show was to feature "three unlikely colleagues—two of them played by de Pablo and Jack Davenport—who inadvertently stumble upon an invention that challenges the very nature of quantum physics—a discovery which in turn puts their lives in grave danger.

[24] On August 28, 2018 Deadline announced that de Pablo and former NCIS castmate Michael Weatherly would be the executive producers of the upcoming CBS detective drama MIA, written by Shepard Boucher; however the series did not proceed to production.