He made his first records with Jay McShann's band in 1945. In 1949, recording under his own name with the McShann band, he had his first hit, "Ain't Nobody's Business", a song which came to be regarded as his signature tune. In 1950 he had hits with two more songs closely identified with him: "No Rollin' Blues" and "Big Fine Girl".
Witherspoon's style of blues became unfashionable in the mid-1950s, but he returned to popularity with his 1959 album, Jimmy Witherspoon at the Monterey Jazz Festival. He later recorded with Ben Webster, Richard "Groove" Holmes, and T-Bone Walker, and toured in the 1970s with a highly regarded band of his own featuring Robben Ford. He continued performing into the 1980s.