The subject of Christ taking leave of his mother derives from devotional, not biblical sources. It relates to the moment when Christ leaves for Jerusalem and anticipates his coming death. The Virgin is shown prostrate with grief, attended by four holy women: Mary Cleophas, Mary Salome, Mary Jacobi and Mary Magdalene. The Virgin's companions also make an emotional appeal to Christ not to leave. Christ is accompanied by Saint Peter and Saint John the Evangelist. The minuscule family of donors at the bottom right have not been identified, although it is probable that they commissioned this picture.
In this, as in other works by Altdorfer, the figures are elongated and their hands and feet enlarged. These distortions emphasise the language of gesture and stance, which Altdorfer uses so effectively. Much of the composition is taken up with landscape, particularly the tall fir trees which Altdorfer often depicted. On the left, through the arch, the sky is red, suggesting sunset and, perhaps, the death to come.