Helen Magnus (Amanda Tapping)
- Must Have Caffeine: Oh, yes, but not coffee. She drinks tea, thank you.Helen: I have standards, Will, and drinking coffee? Far below them.
- Omnidisciplinary Scientist: Justified, since she has had 157 years to study and has been surrounded from birth by the world's leading scientists. Even then, she has mainly focused on the biology and chemistry of Abnormals, rather than any of the social sciences (which is why she hires Will) or the high technology that the Sanctuary uses (which is why she hired Henry), and she seems to only dabble in the physics behind Abnormals.
- Really 700 Years Old: She 157 years old when the series starts but is physically in her 40s. Later she has to take The Slow Path, so it's more like 260, but still no noticeable change in appearance, so it would seem Helen has truly become The Ageless.
Ashley Magnus (Emilie Ullerup)
Will Zimmerman (Robin Dunne)
- Chickification: Will Zimmerman is a rare male example. He's a brilliant forensic investigator, an emotionally tough, tenacious maverick willing to find the truth at any price... in the pilot. In subsequent episodes, he becomes the Designated Victim, losing his glasses and (frequently) 50% of his pilot IQ and personality in the process, as well as gaining tighter clothes and lots of hair product. As of the fourth season premiere, he seems to be getting better.
- Future Badass: In "Pavor Nocturnus". His actions even scare the crap of Magnus of all people.
- Identical Grandson: Captain Jack Zimmerman, Will's identical grandfather, fought in World War II and died saving Helen's life. It's implied to be one of the reasons why Helen hired Will.
- Token Human: Until Kate joined anyways.
Henry Foss (Ryan Robbins)
- Afraid of Blood: "Aw, yeah, come on, doc. You know about me and blood."
- Insistent Terminology: Never call Henry a werewolf. It's hyper-accelerated protean or HAP, thank you very much.
- Our Werewolves Are Different: More experienced werewolves can control their transformations and themselves.
John Druitt (Christopher Heyerdahl)
- Even Evil Has Standards: Druit seemed to be working for the Nazis, though he was really working against them from within, or something (he did kill Hitler after all), because "I know evil when I see it."
- Luke, I Am Your Father: In the pilot it's revealed that he's Ashley's father.
- Really 700 Years Old: Or somewhere in that neighborhood. He's as old as Helen, but probably off by a few years.
- Teleportation Sickness: In the webisodes, this was a particularly dangerous kind that caused his insanity, as it gradually altered his brain chemistry every time he used his power.
The Big Guy
- Bigfoot, Sasquatch and Yeti: He's a Sasquatch.
- Genius Bruiser: The only Sasquatch who can perform surgery.
- No Name Given: He's big and he's a guy. They call him Big Guy.
Kate Freelander (Agam Darshi)
- Action Girl: Her personality didn't change much from her Cabal days, but she's still a pure good guy as shown when she is accidentally Brainwashed and Crazy one episode and still refuses to kill a man who would have been an Asshole Victim. While waiting for Magnus to retrieve the egg of a dangerous abnormal from the vault of a bank, she begins to case the joint apparently as a matter of habit. When the egg hatches and they need to quarantine the bank, her idea is to hold it up in order to prevent the other customers from leaving
- Canada, Eh?: Although one episode established she grew up in Chicago, she moved to Montreal a few years after her father died, and the Sanctuary crew first meet up with her in Vancouver. She notably refers to Will in "Kali Part 1" as "Will Zed," as opposed to "Will Zee" as an American would.
- Jerk with a Heart of Gold: she starts the show with a rather mercenary attitude, but by the beginning of season 3 she views the Sanctuary team as her family, and is particularly affectionate towards Will and Henry
- Put on a Bus: For a good chunk of season 4, then The Bus Came Back.
- Token Minority: She even snarks in an episode where she and Will are in Mumbai that she's "not just the token Indian girl"
- Combat Tentacles: He has an extra appendage at his side.
- The Empath: He is able to sense people's fear. Unfortunately, the more people fear him the more his tentacle has a tendency to attack people
- Really 700 Years Old: Thanks to a self-made vest which keeps him young.
- The Alcoholic: Sort of. As a half-Vampire, alcohol doesn't affect him. He still manages to empty Magnus' wine cellar.
- Game Face: When in vampire mode.
- Hoist by His Own Petard: in de-vamping the leader of the trust-fund vampire kids, he himself is de-vamped, apparently irreversibly. As a consolation prize he does get magnetism powers, however
- Mad Scientist: Well, he is Nikola Tesla.
- The Matchmaker: Will and whoever he might be interested in.
- Omnidisciplinary Scientist: With a bias towards technological invention, although he is shown to be capable in several other fields of science.
- Our Vampires Are Different: And the former rulers of the earth.
- Psycho Electro: He fell asleep in Edison's electric chair!
- Well-Intentioned Extremist: He wanted to create a new batch of vampires so they can seize power around the world and usher in a new age of technological and cultural progress
Walter a.k.a. The Adjuster
- Brought to You by the Letter "S": The suit has a stylized A.
- Clothes Make the Superman: The suit is made out of Abnormal bugs that feed on Walter. As a result, he can't just take the suit off, he has to think about it the right way.
- Cutting the Electronic Leash: In his second appearance, leading up to his He's Back moment.
- They Changed It, Now It Sucks: An in-universe version. He got a movie deal for his superhero persona, but Executive Meddling basically retooled the entire concept. It made him so frustrated that he Took a Level in Jerkass and the suit rejected him when he tried to coax it off Kate. When it eventually did come back to him, he was convinced to go back to his roots and directed the film himself.
- Jekyll & Hyde: The inspiration for the Trope Namer.
- It's Personal: He blames the Five (especially Magnus) for the death of his daughter.
- Never Found the Body: Twice.
- Combat Medic: His prior experience with combat medicine means that he can assist Magnus with the dangerous procedure required to cure Abby in Fugue.
- Hero of Another Story: He's the head of the UK Sanctuary, which is the second largest of them.
- Sixth Ranger: When the team needs an extra hand, they usually call him.
- Stiff Upper Lip: He seems remarkably un-phased by the fact that he is the primary human target of the raid on the UK Sanctuary and thus has six super-Abnormals out for his blood.
- Undying Loyalty: He is a staunch supporter of Magnus.