Barbaro was born in Venice, the son of Zaccaria Barbaro, and the grandson of Francesco Barbaro. At an early age he was sent to Rome, where he studied under Pomponius Laetus. He completed his education at the university of Padua, where he was appointed professor of philosophy in 1477. Two years later he revisited Venice, but returned to Padua when the plague broke out in his native city.
He was sent on various missions to persons of high rank, amongst them Pope Innocent VIII, by whom he was nominated to the important office of patriarch of Aquileia (1491). The Venetian senate, however, refused to ratify the appointment, which, contrary to the law, he had accepted without first obtaining its sanction. He was banished and forced to resign the patriarchate, under the threat of being punished vicariously by the confiscation of his father's property. Barbarus remained at Rome, in receipt of a small pension from the pontifical government, until his death (probably from the plague) in 1493 (according to some, two years later).
Barbaro is remembered for his scholarly work on Aristotle among other writers.