- Legion of Doom
- Royal Flush Gang
- Scarecrow (Mentioned only)
- Blood Tribe (Flashback only)
- Boston Police Department
- Officer in Charge
- Central City Police Department
- Federal Bureau of Investigation
- Agent Porter
- Martha Wayne (Appears only as a corpse)
- Metropolis Police Department (On a TV or computer screen)
- Thomas Wayne (Appears only as a corpse)
- Justice League Watchtower
- United States of America
- Bracelets of Submission
- Flash's Costume Ring
- Green Kryptonite
- Green Lantern Ring
- Justice League Teleporter
- Lasso of Truth
- Scarecrow's Fear Toxin
- Star Sapphire (gem)
Using the Royal Flush gang as a distraction, Vandal Savage has Mirror Master sneak in and steal Batman's files on neutralizing the Justice League. Later Cheetah, Star Sapphire, Metallo, Bane and Ma'alefa'ak arrive in a Louisiana swamp enticed by Vandal Savage's offer of $10 million each. Vandal Savage reveals that he will pay them $100 million plus expenses for each member of the Justice League they eliminate. Using a modified version of Batman's plan they take out the League one by one.
Vandal reveals that his next plan is to use a rocket to trigger a solar flare that will destroy half the planet and disable any technology more advanced then a steam engine. He relates his past and invites Cheetah to test his claim of being immortal; which she does by cutting out his throat. They are all stunned when Vandal get up and the wounds heal before their eyes.
Meanwhile Batman has escaped from the grave he was trapped in and counters the methods used to take out each member of the Justice League. The League then meets and Batman reveals that his files 'call home' if their decryption is broken allowing the League to find the Hall of Doom. They defeat the Legion of Doom but are unable to stop Vandal's rocket. Realizing the Royal Flush were being used to field test intangibility equipment and that the Hall of Doom would have been sun side when the flare hit, the League deduces the Hall of Doom itself is a giant intangibility device. Utilizing their superpowers and the Watchtower, the League used the Hall of Doom to make the entire Earth intangible allowing the solar flare to pass through the planet.
- Kevin Conroy - Batman
- Tim Daly - Superman
- Susan Eisenberg - Wonder Woman
- Nathan Fillion - Green Lantern
- Carl Lumbly - Martian Manhunter, Ma'alefa'ak J'onnz
- Michael Rosenbaum - The Flash
- Bumper Robinson - Cyborg
- Carlos Alazraqui - Bane
- Dee Bradley Baker - Officer in Charge
- Claudia Black - Cheetah
- Paul Blackthorne - Metallo, Henry Ackerson
- Olivia d'Abo - Star Sapphire
- Grey DeLisle - Lois Lane, Queen
- Alexis Denisof - Mirror Master
- Robin Atkin Downes - Alfred Pennyworth, Jack
- Brian George - Mayor
- Danny Jacobs - Special Agent Porter
- David Kaufman - Jimmy Olsen
- Juliet Landau - Ten
- Jim Meskimen - King
- Phil Morris - Vandal Savage
- Andrea Romano - Batcomputer Voice
- Bruce Timm - Ace
- Differences from JLA: Tower of Babel:
- Mirror Master is the one to steal the files from the Batcomputer, not Talia al Ghul.
- Plastic Man and Aquaman are not featured in the film. As well, Barry Allen replaces Wally West as the Flash, and Hal Jordan replaces Kyle Rayner as Green Lantern.
- Cyborg is present in the film, whereas he is absent in the comics.
- At the end of the film, Superman gives Batman the Kryptonite bullet he was shot with in case he is to ever fall from grace. In the comics, it was instead a Kryptonite Ring.
- The tactics to take down the Justice League members is changed from the comics:
- In the film, Martian Manhunter is poisoned with a chemical to make him sweat magnesium. In the comics, his skin is coated with nano machines to turn it into magnesium.
- Green Lantern is exposed to a version of the Scarecrow's Fear Toxin in the film to make him doubt he is worthy of using the Green Lantern Ring. In the comics, he is rendered blind through hypnotic suggestion.
- Wonder Woman is poisoned with a chemical to make every person she sees appear as Cheetah in the film. In the comics, she is injected with a nano machine that would pit her against an evenly matched opponent in virtual reality.
- The Flash has an explosive bracelet bolted through his wrist, set to explode if he were to decelerate, in the film. In the comics, he is shot with a special bullet that caused him to have seizures at light speed.
- Superman is defeated by a Kryptonite bullet in the film. In the comics, he is exposed to a form of Red Kryptonite that caused his skin to become transparent, forcing him to absorb too much solar energy and overload his powers.
- Based on the Tower of Babel storyline from the Justice League of America comics.
- Tim Daly, Kevin Conroy, Susan Eisenberg, Carl Lumbly, Phil Morris, Olivia d'Abo, Alexis Denisof, and David Kaufman reprise their respective roles from the DCAU as Superman, Batman, Wonder Woman, Martian Manhunter, Vandal Savage, Star Sapphire, Mirror Master, and Jimmy Olsen. Although Michael Rosenbaum reprise his role as Flash, this Flash is based on Barry Allen rather than Wally West in Justice League.
- Nathan Fillion reprises the role of Hal Jordan/Green Lantern from Green Lantern: Emerald Knights.
- Bane's statement at the graveyard scene of "When we fought before, I broke the Bat." refers to the Batman storyline "Knightfall".
- Vandal Savage's plan involving solar flares is similar to the Legion of Doom's (although backfired) plan in the episode "History of Doom" of Challenge of the Super Friends.
- The film is in memory of Dwayne McDuffie.
- Characters from Justice League: Doom (Movie)
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- Movie Gallery: Justice League: Doom (Movie)