Dane's first major feature film appearance was in The Basket. He also appeared in Zoe, Duncan, Jack & Jane, Sol Goode, Feast, X-Men: The Last Stand, and starred in Open Water 2. In 2005, Dane guested as Dr. Mark Sloan (nicknamed "McSteamy" by Meredith Grey) in "Yesterday," the eighteenth episode of the second season of the ABC medical drama Grey's Anatomy. Positive audience reaction to the character led to Dane's becoming a regular in the show's third season. His first appearance in the season, in which he walked out of the bathroom soaking wet and wearing only a strategically placed towel, was labeled a "watercooler moment."Dane resigned from the show after the end of season 8 but appeared on the first two episodes of season 9.

In December 2006, he starred in the A&E television film Wedding Wars as the brother of a gay man (played by John Stamos) who goes on strike in support of marriage equality. Dane, alongside Patrick Dempsey, appeared in the same big-screen project, the 2010 romantic comedy Valentine's Day. The Garry Marshall-directed film followed five interconnecting stories about Los Angelenos anticipating (or in some cases dreading) the holiday. In October 2012, Dane joined the main cast of the Michael Bay produced, TNT drama series The Last Ship.

Dane married actress Rebecca Gayheart on October 29, 2004. They have two daughters together: Billie Beatrice and Georgia Geraldine.

Dane made headlines when he participated in a nude tape with his wife and Kari Ann Peniche. Dane checked himself into rehab in June 2011; he entered a California treatment center to rectify his problem with dependency on prescription drugs after suffering a sports injury.

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