He was the elder brother, by about three years, of Hans Holbein the Younger, but he appears to have died in his mid-twenties, leaving behind only a small body of work.

Like his younger brother, he was born in Augsburg (which today is in Bavaria, but then was a free imperial city), a center of art, culture, and trade at that time.

[1] In 1515, Ambrosius is assumed to have lived in the Swiss town of Stein am Rhein, where he helped a painter from Schaffhausen named Thomas Schmid with the murals in the main hall of the St. George monastery.

[4] Franny Moyle writes, "There is no record of Ambrosius's death, but the abrupt cessation of his work in a year [1519] where so many were falling prey to sickness suggests that he either caught the disease that began with a headache, or the plague that came in its wake, and died.

The painting used to be a part of Austrian Imperial connection,[vague] and given the outfit of the young woman is an example of very early 16th century Spanish fashion, closely resembling tomb of Queen Juana I of Castille, it might be an unidentified portrait of her.