Barthel Beham probably learned from his elder brother Hans Sebald Beham and from Albrecht Dürer. During the 1520s Beham was especially active as an engraver, creating tiny technical masterpieces of marvelous detail. He also was interested in antiquity and at some point in his career may have worked with Marcantonio Raimondi in Bologna and Rome.
In 1525, along with the other "godless painters" Hans Sebald Beham and Georg Pencz, Barthel was banished from Lutheran Nuremberg for asserting that he did not believe in baptism, Christ, or transubstantiation. Although soon pardoned, Barthel moved to Catholic Munich to work for the Bavarian dukes William IV and Ludwig X. There his outstanding skill distinguished him as one of Germany's principal portrait painters, sought out by such luminaries as Emperor Charles V. His portrayals were coolly objective, with a strong sense of three-dimensionality.
Beham also was an early creator of vanitas representations, including sleeping children with skulls and women surprised by death. According to Joachim von Sandrart, he died in Italy on a trip sponsored by Duke William.