The show starred Gail Davis as Annie Oakley, with Brad Johnson in the role of Deputy Sheriff Lofty Craig and Jimmy Hawkins as Annie's little brother, Tagg Oakley. Hawkins would ultimately play Tagg in 80 of the series' 81 episodes.

In the series, Annie Oakley rode a horse named Target; Tagg's horse was Pixie; and Lofty's mount was named Forest.  Annie and Tagg lived in the town of Diablo, Arizona, with their uncle, Sheriff Luke MacTavish, who was frequently away whenever trouble started. It would then be up to straight-shooting Annie and her "silent suitor" Lofty Craig to rescue law-abiding neighbors and arrest outlaws. Often Tagg would be told to stay in town and out of the way; but through either disobedience, the need to relay important new information, or being captured by outlaws, he would usually end up in the middle of each episode's adventure.

Annie always wore the same fringed cowgirl outfit, of which 15 or more copies were made throughout the show's production. Her hairstyle was braided pigtails.

Annie Oakley was among Gene Autry's Flying A Productions.