This painting captures a dramatic moment in the apocryphal story of Judith. To save her people, Judith, a Jew, murdered the Assyrian Holofernes, decapitating him when he was drunk. The artist shows Judith with the head of her foe. She glances over her shoulder, the musculature emphasised in Michelangelesque fashion. Drama is lent to the scene through the artist's use of chiaroscuro (extreme contrasts of light and dark) and the diagonal positioning of Holofernes's headless body with the gaping neck in the foreground.

The painting is attributed to an unknown follower of Cherubino Alberti, an Italian artist who worked mainly in Rome where he became Director of the Academy of St Luke.