Sheb Wooley was born in 1921 in Erick, Oklahoma, the third son of william C. Wooley and Ora E. Wooley. He had two older brothers, Logan and Hubert, as well as a younger brother, william. Federal census records for 1930 and 1940 identifiy Sheb's father only as a "Farmer", although the family's livestock holdings apparently included horses, for Sheb learned to ride at an early age and became a working Cowboy and later an accomplished rodeo rider. At the age of 15, with a talent for music, he formed and played in a country-western band, the "Plainview Melody Boys," that periodically performed on radio at station KASA in Elk City, Oklahoma.

Wooley was married four times. His first wife was Melva Miller, whom he married in 1940. Six years later he married Edna Talbott Bunt in Fort Worth, Texas. His third wife was Beverly Irene Addington. He and Beverly remained together for 23 years and adopted one daughter, Chrystie Lynn. Then, in 1985, he married his manager Linda Dotson, who already had a daughter, Shauna.

In the late 1950s, Wooley embarked on a recording career of his own, with one of his hits, "The Purple People Eater." earning him considerable fame. He followed that success with a series of novelty hits, as well as some classic pop recordings and many recordings Classified as Country and Country and Western. In the United Kingdom he enjoyed a minor hit with the comedy single "Luke The Spook" on the flip side of "My Only Treasure," a ballad in the country and western tradition.

He appeared in a 1953 episode of The Lone Ranger titled "Wake of War"; and he performed again on that series the following year in the episode "Message to Fort Apache". He also appeared five times between 1951 and 1955 in the syndicated series The Adventures of Kit Carson. He guest-starred as well in The Cisco Kid in the role of Bill Bronson, portrayed the character Harry Runyon in the episode "The Unmasking" on the CBS Western My Friend Flicka, and appeared twice on the ABC Western series The Life and Legend of Wyatt Earp. Wooley acted too on the "big screen," playing Baxter in the 1958 film Terror in a Texas Town.

In 1954, he played outlaw Jim Younger in the syndicated Western series Stories of the Century. Wooley appeared many times in the syndicated Western series The Range Rider, starring Jock Mahoney and Dick Jones.

Wooley's big break professionally came when he was cast as the drover Pete Nolan in the CBS western Rawhide (1959–1966) with Eric Fleming, Clint Eastwood, and Paul Brinegar. During that time Sheb also wrote and directed some of the episodes. He also continued to act in major films, including Giant, The Outlaw Josey Wales, Silverado, and in Hoosiers starring Gene Hackman, Barbara Hershey, and Dennis Hopper. In the latter film, Wooley portrayed Hackman's longtime friend, Cletus Summers, the principal of Hickory High School.

In the late 1960s and early 1970s, Wooley became a regular on the television series Hee Haw and even wrote the theme song for that long-running series. On Hee Haw he often appeared as the character Ben Colder, playing him as a drunken country Songwriter. Outside of Hee Haw, Wooley even released music and performed as Ben Colder, although he would still sing under his own name as well. Sheb would continue to tour internationally and make personal concert appearances until his death in 2003. Working literally to the end, Wooley recorded his last written song just four days before he died.

Wooley also had a string of country hits, with his "That's My Pa" reaching no. 1 of Billboard's Hot C&W Sides chart in March 1962. That same year, Wooley intended to record the song "Don't Go Near The Indians," but he was delayed by an acting job. Meanwhile, Rex Allen recorded the song, and it was a hit. Wooley, however, would do the sequel to the song, "Don't Go Near the Eskimos," about a boy in Alaska named Ben Colder (had never "been colder"). This sequel was so successful that Wooley continued using the name Ben Colder, with one of his later recordings being "Shaky Breaky Car" (which parodied the song "Achy Breaky Heart"). In December 1963, his single "Hootenanny Hoot" became a top-10 hit in Australia; and in 1967 his song "The Love-in" (1967) was recognized as an acerbic commentary on the 1960s' counterculture.

Wooley was the recipient of numerous awards over the years for his accomplishments as a singer, an actor, and as a Writer for both comedic and dramatic productions. In 1968 he received the Country Music Association's Comedian of the Year Award. He also received the 1992 Songwriter of the Year, two Golden Boot Awards, and he won the Western Heritage Award for nine consecutive years in recognition of his film and television work in Westerns.

Wooley was diagnosed with leukemia in 1996. After seven years of battling the illness, he died at the age of 82 at the Skyline Medical Center in Nashville, Tennessee, on September 16, 2003. He was entombed in Hendersonville Memory Gardens in Hendersonville, Tennessee. His illness had forced him to retire from public performing in 1999.