The museum was founded in 1934, as the heir of the Pinacoteca Regia and the Galleria Reale, which had been established in Palazzo Madama by King Charles Albert of Savoy in 1832.

A Civic Museum had been founded in 1860 in the wake of the unification of Italy although, three years later, the collections were moved to another location in Turin, in Via Gaudenzio Ferrari.

The 15th-century Torre dei Tesori ('Tower of the Treasures') is home to several of the museum most known works: Antonello da Messina's Trivulzio Portrait, the Turin–Milan Hours, and several objects from Charles Emmanuel I's cabinet.

Other works include a series of sculptures of the Dead Christ, paintings by Macrino d'Alba, Giacomo Jaquerio, Giovanni Martino Spanzotti, Defendente Ferrari, Antonio Vivarini, Giulio Campi, Gaudenzio Ferrari, Gandolfino da Roreto, Gerolamo Giovenone, Francesco Hayez, sculptors' and goldsmiths' works from the 8th to the 13th centuries, and Piedmontese coats of arms.

The Camera delle Guardie ('Guards Chambers') houses Baroque paintings by artists such as Orazio Gentileschi (Assumption and St. Jerome), Giovanni Battista Crespi, Giulio Cesare Procaccini and Francesco Cairo.