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John Winchester – Jeffrey Dean Morgan

John was a US Marine, presumably until he met his wife, Mary, and settled down in Lawrence, Kansas with her, having two boys, Sam and Dean. In Lawrence, he co-owned a garage with his friend Mike. After Mary's death, he visited a psychic named Missouri. She showed him what was out there in the world and he realized that what he'd seen the night of Mary's death could be explained, even if no-one wanted to believe him. He brought the boys up by himself, all the while searching for answers about what exactly it was that killed his wife. He learnt as much as he could about the supernatural and trained himself and his sons to fight evil in whatever form it took.

When Sam left home to go to college, John continued to work with Dean, until he went missing while on the trail of a White Woman in Jericho, California. He left Dean a voicemail message warning him to be careful. Dean and Sam searched for John for months with no sign of him. Little did they know that when they went home, he was there, hiding out in Missouri's house. Later he phoned the boys from a payphone in Sacremento, California. He was not heard from until Sam and Dean believed they had found the demon they were all looking for in Chicago. It was a trap, however, and all three were nearly killed by a Daeva. John parted ways from the boys again in order to protect them.

When they finally got back together, it was with a greater chance of defeating the Demon. An old friend of John's left him a mystical gun that would be able to destroy the Demon. The three of them faced the Demon, but tragedy struck when their car was smashed into by a truck whose driver was being possessed by the Demon. John and Sam survived with little wrong with them, but Dean wound up in a coma. When John realised his eldest son was going to die, he made a deal with the Demon - offering the gun and his own life in exchange for Dean's. Minutes later, Sam found John laying dead on the floor.

The boys cremated John's body in a pyre, taking his US Marine dogtags to Mary's headstone so that the two of them could share something.
Mary Winchester (born 1954) is the wife of John Winchester, and mother of Dean and Sam. She gave birth to Dean on January 24, 1979 and Sam on May 2, 1983.

In the episode "Pilot," it is revealed that on November 2, 1983, Mary woke after putting her six-month-old son, Sam, to bed. She entered his nursery to find someone she thought to be John leaning over his crib. Returning to bed, she noticed a television flickering downstairs. She found her husband John asleep in the living room, and ran back to Sam's nursery. Hearing his wife's screams, John ran into the nursery. In Sam's room, everything appeared normal, until he saw droplets of blood next to Sam's head. Looking up, he found Mary pinned to the ceiling with a large gash through her stomach. She then burst into flames, dying at the age of 29. She was murdered by Azazel.

In 2005, her sons returned to their childhood home to investigate a recurring dream Sam had experienced in the episode "Home." Mary's spirit seemed to haunt the house in Lawrence, Kansas, and a poltergeist was in the house with her, though the psychic Missouri Moseley never felt their presence. After the poltergeist pinned Sam to the wall, Mary appeared as a fiery figure. When Dean broke through the locked door to reach him, Mary told Sam that she was sorry. Turning to the ceiling, she told the poltergeist to leave her sons alone. Her spirit fought with and canceled out the poltergeist, forcing both spirits out of the home.

In All Hell Breaks Loose, Part 1, the Demon shows Sam the night Mary was killed because she was in the wrong place at the wrong time. When Mary ran back into the nursery she said "It's You!" to the Demon. It remains unclear how she could have known the Demon. In Supernatural: Origins, it seems that Mary has a sister that the Demon was apparently after, but this is unconfirmed. In The Kids are Alright we learn that all of Mary's friends and acquaintences have been killed off over the years since her death.

Mary Winchester – Samantha Smith

Jessica Moore - Adrianne Palicki

Jessica Lee Moore was the girlfriend of Sam Winchester. She was killed by Azazel because she got too close to Sam, who Azazel hoped would be a soldier in his war. She appeared once as an apparition to Sam, but it is unclear if she was just a hallucination. Jessica's death is what drove Sam to be so dedicated to killing Azazel.
Azazel has several children of his own, notably his daughter, the demon who possessed Meg Masters, an antagonist in the first season who was exorcised and sent back to hell by Sam Winchester. The only thing he seems to fear is The Colt, an enchanted gun which can kill anything. He spent the first season trying to obtain this weapon. At the beginning of the second season, he procured it from John Winchester, along with John's life, in exchange for saving the life of John's son, Dean.

Azazel / Yellow-Eyed Demon – Fredric Lehne

Azazel’s son - Sebastian Spence

Azazel’s son has only appeared in two episodes, Salvation and The Devil’s Trap. He was pronounced Azazel’s son in The Devil’s Trap after Dean killed him with the Colt. Not much else is known about him.
: Gordon Walker was 18 years old when a vampire broke into his house and attacked his sister. Gordon grabbed his father’s shotgun, attempted to free his sister, and then finally shot at the vampire. The vampire threw him across the room, knocking him unconscious, and abducted his sister. When Gordon regained consciousness, the vampire and his sister were gone.Gordon ran away from home learning to fight, hunt, and kill vampires. He eventually found the vampire that had taken his sister and killed it. He also killed his sister, as she was then a vampire. This marked the beginning of his hatred for the undead. He became enemies with the Winchester boys when he tried to kill vampires that weren’t hurting people. He then discovered the rumors about Sam being the Anti-Christ and tried to kill him and Dean multiple times. Sam eventually killed him in Fresh Blood after a vampire had turned Gordon.

Gordon Walker - Sterling K. Brown

Kubrick - Michael Massee

Also a hunter, who hunted with Gordon Walker, who was also convinced that Sam is the Anti-Christ. He was killed by Gordon.
Ash attended MIT which according to Ash is "a school in Boston", before he was kicked out for fighting. He sported a haircut which is "all business in front, party in the back". It is unknown when he met Ellen and Jo Harvelle, but he lived in the Roadhouse Saloon, which Ellen owns. He had a room in the Roadhouse with a sign on the door reading, "Dr. Badass is" with the option of "in" or "out". He owned a homemade laptop, which he used to track the paranormal, particularly The Demon, with the information that John Winchester and his sons, Sam and Dean, have gathered. In the episode All Hell Breaks Loose, Part 1, he called Dean to tell him something, but once Dean arrives at the Roadhouse he finds someone to be wearing the same watch as Ash, burned along with the Roadhouse. In All Hell Breaks Loose, Part 2, Ellen informs Sam, Dean, and Bobby that Ash died in the explosion at the Roadhouse, after urging Ellen to go check the safe in the basement for a vital piece of information which ultimately helped them find and confront the Demon.

Ash - Chad Lindberg

Jake Talley – Aldis Hodge

One of the Special children, Jake had powers of predicting the future and super strength. He killed Ava and Sam, but when Sam was brought back to life, he killed Jake.
One of the Special children, Ava had the psychic powers of predicting the future. Her fiancé was killed when she was kidnapped by Azazel, but it is unclear whether she killed him or if he was killed by another demon. She later gained the power of controlling demons. She is killed by Jake Talley when she attempts to kill Sam after turning evil.

Ava Wilson - Katharine Isabelle

Andrew “Andy” Gallagher - Gabriel Tigerman

One of the Special children with powers of mind control. He killed another of the special children, Ansem Weems, before Ansem could kill an innocent girl. He was killed by Ava Wilson.

Agent Hendrickson appeared in three episodes, Nightshifter, Folsom Prison Blues, and Jus in Bello. In Nightshifter he announces his presence to Dean obnoxiously over the phone as Dean is in a bank ‘holding hostages’ and it is realized that he is building a case against Dean and Sam. In Folsom Prison Blues, he finally meets Dean and, after some banter, leaves when he lawyer comes in. He is convinced that Dean is a monster (though not the literal kind that Supernatural fans are familiar with) since he thinks Dean's murdered many people. In Jus in Bello he becomes possessed by a demon and, after Sam exorcises him, he finally believes the truth about the monsters out there. He assists in the fight against a demon horde that descends on the jail the boys are in, but is killed in the last minutes in the episode by Lilith the demon.

FBI Agent Hendrickson - Charles Malik Whitfield

Bela Talbot – Lauren Cohan

Presumed dead; death by hellhounds. Described by Bobby Singer as "pretty friggin' far from a hunter", Bela is an attractive Englishwoman who acquires magical objects for wealthy clients. Bela is amoral, claiming that she will sell anything to anyone for the right price. She only shows an ability to help Sam and Dean when it benefits her. Dean threatens to kill Bela numerous times. It was implied that she once killed one of her own family members.