His film work includes roles in 28 Days (2000), A Knight's Tale (2001), Dodgeball: A True Underdog Story (2004), voice and motion capture for Sonny in I, Robot (2004), and 3:10 to Yuma (2007).

While in college, he played Beaver Smith in an eastern New Mexico summer stock theater production of Billy the Kid.

Tudyk said of the role, that a doctor had given him a shot of adrenaline prior to filming as he had an allergic reaction to food the night before and his windpipe was closing up.

In 2005, he reprised his role as the playful, easy-going Hoban "Wash" Washburne in the film Serenity, derived from the television series Firefly by Joss Whedon.

[16] He voiced the Duke of Weselton in Frozen, Alistair Krei in Big Hero 6, Duke Weaselton in Zootopia,[17] Heihei in Moana, KnowsMore in the Wreck-It Ralph sequel Ralph Breaks the Internet, Tuk Tuk in Raya and the Last Dragon,[18] Pico the toucan in Encanto,[19] Duffle in Strange World, and Valentino in Wish.

Additionally, in 2016, Tudyk played the droid K-2SO in Rogue One: A Star Wars Story, and provided the voice of the parrot, Iago in the 2019 live-action adaptation of Aladdin.

[20] Tudyk also voiced the Mad Hatter in the 2023 short film Once Upon a Studio, made to celebrate the centennial of The Walt Disney Company.

[23] Tudyk filled in for Hank Azaria's roles in Spamalot from June to December 2005, and starred in a limited run of Prelude to a Kiss.

Tudyk portrayed the playful, easy-going Hoban "Wash" Washburne in the 2002 television series Firefly by Joss Whedon.

[12][4] The series ran for one season, and Universal Studios purchased the rights to make a film, Serenity, in which Tudyk reprised his role.

Among several guest spots on shows such as Arrested Development, he played a convicted child sex offender on an episode of CSI: Crime Scene Investigation.

Tudyk also guest-starred in three episodes of ABC's modern remake of the television miniseries V. He provided the voice of superhero Green Arrow and villain Psimon in the animated series Young Justice.

He also provided the voices of Ludo Avarius and King River Butterfly in the Disney animated series Star vs the Forces of Evil.

In 2015, Tudyk released his own web series Con Man based loosely upon his experiences touring the convention circuit after the cancellation of Firefly.