Luini was born as Bernardino de Scapis in Runo, a frazione of Dumenza, near Lake Maggiore. Details of his life are scant.
In 1500 he moved to Milan with his father. According to Lomazzo, he trained under Giovan Stefano Scotto, although for others he was a pupil of Ambrogio Bergognone. In 1504-1507 he was probably in Treviso, as attested by a Madonna with Child signed Bernardinus Mediolanensis faciebat which is however of disputed attribution. His first fresco works are an Adoration of the Magi in San Pietro of Luino (c. 1505) and the attributed fresco in the presbytery of Monza Cathedral with St. Gerard of the Painters.
Luini returned to Milan in 1509, receiving a commission for a polyptych from which today only St. Anthony of Padua in the Museo Poldi Pezzoli, influenced by Bernardino Zenale's Cantù Polyptych remains. In the 1510s he painted frescoes in the Oratory of Santa Maria Nuova in Pilastrello, a Lamentation of the dead Christ in Santa Maria della Passione, a Madonna della Buonanotte in the Abbey of Chiaravalle, frescoes in San Giorgio di Palazzo (1516) and in the Certosa di Pavia, and others.

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