The Virginian is a tall, black-haired cowboy with a deep Southern drawl. He works as a hired hand at Sunk Creek Ranch, where his intelligence and competence quickly earn him the role of deputy foreman. He holds a strict personal code of honor and approaches both his work and his courtship of the local schoolteacher with patient determination. Despite his rough exterior, he seeks to improve his education and understands the value of civility.

Molly is a twenty-year-old schoolteacher who leaves her comfortable Eastern life to work in the Wyoming territory. She descends from the Revolutionary War heroine Mary Stark and shares her ancestor's fiercely independent spirit. She values her career and resists conforming to the expectations of her wealthy relatives. Though she is drawn to the frontier, she maintains strict standards of behavior and wrestles with her attraction to a man outside her social class.

The Narrator is a young, wealthy man spending his summer at Sunk Creek Ranch. Earning the nickname "the tenderfoot," he frequently struggles to understand the harsh realities of frontier life and relies on the cowboys to keep him safe. He observes the locals closely and deeply admires the raw capabilities of his Western friends. He serves as a willing student of the territory and the people who tame it.

Trampas is a rough-edged cowboy who resents authority and discipline. Following a tense standoff over a card game, he harbors a bitter grudge against the Virginian. He actively seeks ways to undermine his rival while plotting his own schemes among the ranch hands. He uses intimidation and manipulation to influence weaker men to join his illicit activities.

Balaam is a nearby rancher known for his severe temper and harsh methods. He believes animals must be physically broken to obey orders, regularly beating his horses. His cruel behavior puts him at odds with cowboys who prefer a gentler handling of their livestock. He prioritizes his own convenience over the well-being of the creatures in his care.

Shorty is a small, somewhat dim-witted cowhand who works hard but struggles to save his earnings. Despite his flaws, he demonstrates genuine kindness toward his horses and possesses a good heart. He easily falls under the influence of manipulative men, ignoring the advice of those who genuinely want to help him.

Steve is an old friend of the Virginian from their early days in Medicine Bow. He enjoys mischievous pranks and the rowdy camaraderie of town life. He works as a cowboy but displays an increasingly relaxed attitude toward the law, a shift that threatens his standing with his more principled friends.

Judge Henry is the wealthy owner of Sunk Creek Ranch. He is a wise and pragmatic employer who recognizes the talents of his men, quickly promoting those who show initiative. He believes deeply in frontier justice and serves as a mentor to the younger generation working on his land.

Mrs. Henry is the wife of Judge Henry. She helps bring the comforts of Eastern civilization to the Wyoming territory, providing a more refined environment at the ranch. She actively assists the cowboys with their education, correcting their spelling and penmanship.

Mrs. Taylor is a practical frontier woman who lives next door to the schoolhouse. She befriends the new schoolteacher and offers blunt advice regarding romance and Eastern pride. She actively supports Molly's integration into Western society and steps in to help during medical emergencies.

Mr. Taylor is a local resident of Bear Creek. He helps facilitate introductions between the established cowboys and the incoming Easterners. He is a reliable neighbor who assists his community when trouble strikes.

The Great-Aunt is Molly's elderly relative. Having lived a long life, she possesses a broad perspective on romance and independence. She understands Molly's desires far better than the rest of their conservative family and encourages her niece to follow her heart.

Scipio is a lithe, talkative cattleman. He takes a job as a cook for the Sunk Creek Ranch crew after a chance meeting on the railway. He quickly recognizes the Virginian's intelligence and becomes a loyal ally, offering keen observations about the social dynamics among the cowboys.

Sam is a wealthy and respectable young man from Molly's hometown. He actively pursues her hand in marriage, representing the safe, conventional life she leaves behind. His predictable nature fails to capture Molly's imagination.

Dr. MacBride is a severe preacher visiting the Wyoming territory. He delivers harsh sermons focused heavily on damnation, with little room for forgiveness. His rigid, humorless demeanor makes him a prime target for the cowboys' practical jokes.

Lin is a local cowboy who previously attempted to court Molly Wood but accepts her rejection gracefully. He participates in playful mischief with his fellow ranch hands, including a notorious baby-switching prank. He maintains a good-natured attitude toward his peers.

Pedro is an excellent pony belonging to Shorty. He receives kind treatment from his original owner but faces brutal conditions when sold to settle debts. His experiences reflect the harsh reality faced by livestock on the frontier.

Uncle Hughey is an older resident of the territory who frequently travels in search of romance. The younger cowboys playfully tease him about his marital ambitions, though he generally enjoys the attention. He eventually returns to town with a new, younger bride.