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Voiced by: Jerry Trainor
- Catch-Phrase: "Hi-gee-gee!"
- Disappeared Dad: There's never any mention of his dad and seeing how his mom briefly dated Snaptrap, Dudley's dad is either divorced from Peg or dead.
- Funny Animal: Dudley is a walking, talking secret agent, but still harbors affection for chewing his own butt and chew toys.
- Genius Ditz: Dudley, who seems to be little more than a hyperactive Idiot Hero, has flashes of genuine cleverness and defeats villains through outsmarting them almost as often as he out-muscles them.
- Half-Dressed Cartoon Animal: Averted in "Mom-A-Geddon" where it's shown that Dudley wears underpants underneath pants that look like naked animal legs.
- LEGO Genetics: Dudley is a combination of every breed of dog ever, and somehow a bit of goat. Partially justified as a long enough blood line could mix every dog breed, the goat is something else.
- Lethally Stupid: His stupidity is the cause of most of Kitty's injuries. And it's not just Kitty who's a victim of these. Very few characters are safe around Dudley.
- Man Child: He lives in a multi-story tree house near his mom's house, and joins in with the kids when Snaptrap uses the "no hitting or killing near kids" rule to his advantage, doing kid related stuff along with crimes.
- Tiny Guy, Huge Girl: With Kitty.
- Too Dumb to Live: He doesn't even know what a toaster does.
- Forget toaster. He's doesn't even know how a light switch works.
- Would Hit a Girl: Dudley doesn't have any problem hitting Kitty, even kicking her in the very same episode, also being seen giving her slaps, bites and, what could be, punches.
Voiced by: Grey DeLisle
- Aloof Dark-Haired Girl: Being the most composed and aloof one in the main cast comes with being the Only Sane Woman.
- Attention Deficit... Ooh, Shiny!: In spite of all her time lecturing Dudley about this, Kitty is easily distracted by cat stuff: mice, scratching posts, catnip, etc.
- Berserk Button: Never pull Kitty's tail, as the Chameleon learned the hard way in "Bark to the Future".
- A Dog Named "Dog": A cat named Kitty.
- Cain and Abel: The Abel to Katty's Cain.
- Cats Are Superior: With Kitty, it should probably come as no surprise, as she is naturally the brains of the duo.
- The Chew Toy: Takes even more Amusing Injuries than Dudley.
- Cute Kitten: Kitty as a kitten is just darn adorable.
- Expy: Has characteristics matching Catwoman especially in Batman: Arkham City and Batman: Arkham Origins Blackgate.
- Funny Animal: Kitty has little to no balance when her whiskers are cut, as the popular (but incorrect) myth states about real cats.
- Informed Deformity: In "Diary of a Mad Cat" there's a Running Gag where people point out Kitty's "broad shoulders."
- Ms. Fanservice: She gets a lot of moments where her curves are shown off.
- The One Who Wears Shoes: In her case, they're Combat Stilettos.
- Slapstick Knows No Gender: Thanks to her Chew Toy status.
- The Smurfette Principle: Kitty is the only female in the main cast.
- Species Surname: Kitty Katswell.
Voiced by: Jeff Bennett
- Cartoon Creature: Word of God says, "[T.U.F.F.] always knows [Keswick] will have the answer, but they just don't know what kind of animal he is." Its revealed in a later episode that Keswick is actually part of his own species, also called Keswick. They have quite an unusual life cycle, including a 'platypus phase' where they shoot toxic quills when they hiccup.
- Gadgeteer Genius: He's made a machine that makes donuts out of thin air and redesigned D.O.O.M.'s lair using soup cans and a shark tank.
- Last of His Kind: Possibly, since he is part of his own species. Part of their life cycle is growing a reptilian tail that breaks off and becomes another Keswick, who they must fight to the death.Keswick: There can be only ONE Keswick!
- LEGO Genetics: Keswick has gills, webbed feet, lays eggs, has a pouch, and who knows what else.
Herbert "The Chief" Dumbrowski
Voiced by: Daran Norris
- Artificial Limb: The Chief has a bionic foot that makes him even cooler than he is already.
- Pint-Sized Powerhouse: He's extremely strong for somebody so small.
- Pointy-Haired Boss: Could qualify. Although he has extreme strength for his size and is good in a direct fight with villains, he doesn't seem to actually have the skills needed to lead a spy organization.
- Real Men Wear Pink: In "The Rat Pack", it's revealed that he reads romance novels, though he tries to deny it and say it's hard-boiled detective fiction.
- Retired Badass: The Chief was once T.U.F.F.'s top agent and an amazing fighter. In one episode, he comes out of retirement to help Dudley deal with the Chameleon. While things don't work out well at first, Dudley manages to give him a pep talk and he shows he's still got it by beating up the Chameleon.
Voiced by: Matthew W. Taylor
- Catch-Phrase: "It is I, Verminious Snaptrap!" and "I shall now plot my revenge!" He also often ends conversations with "Snaptrap out!"
- "Start the car Larry!" whenever committing an act of petty theft.
- Deadpan Snarker: Has his moments. Especially when Larry in involved.
- Even Evil Has Standards: Snaptrap would never kill anybody on their birthday without letting them celebrate it first. "No one's THAT evil!"
- When Snaptrap asks for suggestions on how to get revenge on the "Meanies" at the monotrail for not letting them ride, Francisco suggests that they eat them. Snaptrap flat-out states it's just "dark and disturbing".
- The reason why Snaptrap tells the agents his latest plan is because he considers it rude not to.
- Evil Is Petty: In addition to doing things that qualify as evil, most of Snaptrap's actions seem to Poke the Poodle.
- Freudian Excuse: Might qualify considering all the hints dropped on the show about his mother being abusive, and how much he hates her.
- Laughably Evil: Some of his schemes are so pathetic it's funny.
- No Celebrities Were Harmed: Snaptrap's voice is an impersonation of Ed Wynn.
- Psychopathic Manchild: Most of his schemes are very petty and childish. For example, he tried to blow up the sun just to make popcorn in one episode.
- Rodents of Unusual Size: He's a rat as tall as Agents Puppy and Katswell.
- Speech Impediment: Has a lisp.
- Token Evil Teammate: During his brief tenure as a T.U.F.F. Agent in "Snap Dad".
- Too Dumb to Live: Is a mild example, particularly in "Mission: Really Big Mission" where his get rich quick scheme for the episode will also destroy the entire planet. Even The Chameleon calls him out on his embarrassing case of Didn't Think This Through.
- Vocal Evolution: His voice was initially a bit deeper, but cracked constantly - but by the end of the first season his voice became more consistently high-pitched, lessening the Ed Wynn aspect of his voice.
Voiced by: Daran Norris
- Attention Deficit... Ooh, Shiny!: Has a hard time focusing on his schemes over his stoma- OOH LADYBUG! *nom*
- Catch-Phrase: "It is I, The Chameleon!"
- Freudian Excuse: He's a villain due to being a social outcast.
Voiced by: Rob Paulsen
- Animal Gender-Bender: He has laid eggs for comedic effect.
- Duck!: A running gag with many of his minions, including a duck named duck (and Skip). In "Bad Eggs", he shows an application form for being one of his henchmen, which includes a space for "weird name".
- Last of His Kind: He uses this to his advantage to protect himself from T.U.F.F. Until he violates this by creating a giant mutant boobie, therefore no longer protected by the law for being the last one, despite the fact that he'll still be protected for being an endangered species.
- Only Sane Man: When you compare him to Snaptrap and The Chameleon.
- Rail Enthusiast: He really likes taking the bus and he even memorized different bus schedules. Only because he can't fly.
- Surrounded by Idiots: Not only when dealing with his henchmen, but when he teams up with Snaptrap and The Chameleon.
- Younger Than They Look: According to "A Doomed Christmas" he's 23, though that doesn't stop everyone else from thinking otherwise.
Quacky the Duck
Voiced by: Matthew W. Taylor
- Excited Kids' Show Host: He even continues to act like this after he gets arrested, though he's considerably calmer.
- Early-Bird Cameo: Is mentioned and shows up on television a few times before he becomes a supervillain.
- Face–Heel Turn: He turns evil after his show gets cancelled by the network president.
- Falsely Reformed Villain: Most of his latter plans include pretending to go straight and open some kind of business.
- Genre Savvy: Is entirely aware that Dudley's an idiot, and often uses it to his advantage.
- No Celebrities Were Harmed: Quacky's voice is an impersonation of Bobcat Goldthwait.
- Paper-Thin Disguise: When attempting to disguise himself as someone else, he does an extremely horrible job at hiding who he is. T.U.F.F. believes him anyway.
- Serious Business: His Show Within a Show is insanely popular. Even after he gets arrested, he's so famous he's able to become a Villain with Good Publicity with almost no effort.
- Suspiciously Specific Denial: A running gag. He's pretty bad at hiding his plans. Lucky for him, Dudley's a moron.
The Sharing Moose
Voiced by: Jeff Bennett
Voiced by: Leslie Carrara-Rudolph
- Parent with New Paramour: In "Snap Dad".