One of the most influential artists in sixteenth-century Florentine painting, Santi di Tito was a key figure in the transition between Mannerism and the Baroque. 

Santi undertook training in Florence under Agnolo Bronzino and then with Baccio Bandinelli. During this time he experimented with various elements that were to define his style throughout his mature career. Between 1558 and 1564, Santi was working in Rome where he absorbed the classicism of Raphael. On returning to Florence in 1564, Santi joined the Accademia del Disegno and began working with Vasari on court commissions, fully embracing the Mannerist style. Over the next ten years, Santi produced paintings that demonstrate a knowledge of various artists and schools; following a visit to Venice in 1571-72, the influence of Paolo Veronese is evident in Santi's Feast in the House of Simon and the realism of the figures in his Vision of St Thomas Aquinas (1573) reflect a familiarity with Lombard painting. 

Following Vasari’s death in 1574, Santi received fewer court commissions and worked increasingly for churches, confraternities, and private patrons. Stylistically he returned to the classicism of Raphael, and his religious works in particular are distinguished by their narrative clarity and sincere religious sentiment, tempered by a gentle naturalism, all of which were in line with Counter-Reformation preferences. His production of highly-finished drawings which were then squared for transfer is indicative of the Mannerist tradition, but Santi's desire to achieve greater naturalism in his figures led to him combining this technique with the influences of Andrea del Sarto and Raphael. This fusion of styles catalysed a reform of Florentine painting.

In the latter stages of his career, Santi took a greater interest in the use of light and colour that was being introduced by younger Florentine artists such as Jacopo da Empoli and Lodovico Cigoli, many of whom were working in his workshop. He was also a well-respected teacher and when, in 1602, the Accademia del Disegno established rules for the protection of Florence's artistic patrimony, he was mentioned as a leading artist whose signature was required to sanction the exportation of artworks.

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