The gameplay has been described as action-heavy, and compared to the Ratchet & Clank series. Throughout 24 levels in total, players enter sci-fi vehicles and blast enemies with guns, encounter 6 boss fights, and have other gaming experiences. GameSpot described it as having "Smash TV-esque mechanics".
Inspired by the "superhero movie within the movie" finale of the movie, Chicken Little: Ace in Action features Ace, the superhero alter ego of Chicken Little, and the Hollywood versions of his misfit band of friends: Runt, Abby and Fish-Out-of-Water. The crew of the intergalactic Battle Barn faces off against Foxy Loxy and her evil Amazonian sidekick, Goosey Loosey, who have an evil plan to take over Earth. Battle evil alien robots through multiple levels across the solar system and combat your foes in one of three distinct game play modes: Ace on foot as a soldier, Runt as the driver of an armored tank, or Abby as the pilot of a spaceship. The original Chicken Little and his friends Abby, Runt and Fish we know from the film are featured in cut scenes throughout the game.