Main Autobots:
* Optimus Prime: Peter Cullen
* Bumblebee: Mark Ryan (voice)
* Ironhide: Jess Harnell (voice)
* Ratchet: Jeffrey Combs (voice)
* Jolt: Charlie Adler (voice)
* Sideswipe: Roger Craig Smith (voice)
* Skids: Tom Kenny (voice)
* Mudflap: Reno Wilson (voice)
* Arcee: Lisa Ann Beley (voice)
* Jazz: Darius McCrary (voice, appears only in flashbacks)
* Wheelie: Tom Kenny (voice)
Other Autobots:
* Sentinel Prime: Leonard Nimoy (voice)
* The Wreckers: A team of Autobot warriors including Topspin (Kevin Michael Richardson) and Roadbuster (Reno Wilson)
* The Constructicons: A team of Autobot warriors including Scavenger, Mixmaster, Long Haul, Scrapper, and Hook (various voice actors)
Note: This list is not exhaustive, as there are other Autobots that appear in the film for brief periods.