Wooley was born in Erick, Oklahoma and grew up on a farm. He learned how to ride horses at a young age, and was a working cowboy and rodeo rider. He also played in a country-western band. During World War II, Wooley was turned down for service because of his rodeo injuries. He worked in the oil industry and as a welder. In 1946, he moved to Fort Worth, Texas and became a country and western musician.
Wooley appeared in dozens of western films from the 1950s through 1970s, most notably High Noon. He also appeared in The Outlaw Josey Wales and Giant. He also co-starred as Pete Nolan in the TV Western Rawhide.
In the late 1950s, he embarked on a recording career, and recorded the song that made him famous. Wooley followed up "People Eater" with a series of lesser-known novelty hits. Wooley also wrote the theme song for the long-running television show Hee Haw.
Following his success with "Purple People Eater," Wooley enjoyed a string of country hits, his most successful being "That's My Pa," which reached No. 1 of ''Billboard magazine's Hot C&W Sides chart in March 1962.
He was a regular on Hee Haw as the drunken country songwriter Ben Colder. The Colder persona became popular and he released music and performed under that name as well as his own.
The Ben Colder persona was created after an incident in which Sheb Wooley was supposed to record the song "Don't Go Near The Indians", but was delayed due to an acting job. During the delay Rex Allen recorded the song and scored a hit, and so Sheb Wooley told people that he didn't mind - he would do the sequel. His version was "Don't Go Near the Eskimos", about a boy who lives in Alaska, and as an extra joke he used the name Ben Colder (as in living in Alaska means he had never "been colder"). The single was so successful he continued using the persona for another forty years, with one of his last recordings being "Shaky Breaky Car" (which parodies the song "Achy Breaky Heart").
He is considered by many to be the most likely voice actor for the Wilhelm scream, having appeared on a memo as a voice extra for Distant Drums. This particular recording of a scream has been used by sound effects teams in over 130 films.
Wooley continued occasional television and film appearances through the 1990s, including a notable appearance as "Cletus", the principal in the 1986 film Hoosiers. In 1996 he was diagnosed with leukemia, and died at the Skyline Medical Center in Nashville, Tennessee in 2003.