The dance act will headline a set at London's Roundhouse on September 9 as part of the music service's month-long series of free-ticketed gigs.
Deadmau5 joins a lineup which already includes Usher, One Direction, Rebecca Ferguson, Olly Murs, Jack White, Emeli Sandé and Ed Sheeran.
Fans can apply for tickets now through the iTunes Festival website or accompanying iOS app.
The DJ is currently working on the follow-up to his 2010 album 4x4=12, which is expected to be released later this year.
Meanwhile, Deadmau5 has teamed up with My Chemical Romance frontman Gerard Way for new track 'Professional Griefers', which will be available to download from September 2.
Listen to Deadmau5 and Gerard Way's 'Professional Griefers' below: