Margaret of Austria (1480-1530; also depicted in RCIN 403428) was the aunt of the Holy Roman Emperor Charles V and acted during his infancy as Regent of the Netherlands from 1506-15. She ruled the Netherlands again as Governor (reporting to Charles as King Charles I of Spain) from 1519-30.

She is depicted facing the spectator with her head turned half to the left. She wears a white coif and barbe and a black dress edged with brown fur and rests her hands on a green table top.

Bernard van Orley lived and worked mainly in Brussels. He was the official court artist to Margaret of Austria from 1518 until her death in 1530, and he is known to have painted at least twelve portraits of Margaret of Austria. Van Orley ran a large workshop which produced numerous portraits and religious paintings, as well as designs for tapestries and stained glass.