The gallery houses the most complete collection of Umbrian artworks dating to between the 13th and 19th centuries, most of which came from ecclesiastical properties. However, the close relationship between the religious and civil ambits throughout the Middle Ages and for most of the Modern era ensures that this museum is one of the best repositories of the artistic heritage not only of the region of Umbria but of the country as a whole. 

The artistic movements of the 17th and 18th centuries are represented by artists such as Orazio Gentileschi, Pietro da Cortona, Valentin de Boulogne, Pierre Subleyras, Antonio Amorosi and Corrado Giaquinto. The Gallery also features various special collections, such as textiles, documents and maps, and a Treasury with precious art- works from other city museums and churches.


Worthy of mention are the works by some of the major artists of the Middle Ages and the Renaissance, such as the Master of San Francesco, Arnolfo di Cambio, Duccio di Buoninsegna, Gentile da Fabriano, Beato Angelico, Piero della Francesca. There are many masterpieces by Pietro Vannucci, better known as Perugino, and by his followers and other local artists, the most important of which is Bernardino di Betto, called Pintoricchio. Interesting is the Cappella dei Priori, a chapel frescoed by Benedetto Bonfigli, with scenes from the life of Saint Ercolano and Saint Louis of Toulouse, an example of local history and town planning.