Originally a nine-bedroom Georgian townhouse built in the 1830s on the footpath leading to Kilburn Abbey, the building was later converted to apartments where the most flamboyant resident was Maundy Gregory. The premises were acquired by the Gramophone Company in 1931 and converted into studios. Pathé filmed the opening of the studios, when Sir Edward Elgar conducted the London Symphony Orchestra in recording sessions of his music. The neighbouring house is also owned by the studio and used to house musicians. During the mid-1900s the studio was extensively used by leading British conductor Sir Malcolm Sargent, whose house was just around the corner from the studio building.

The Gramophone Company later amalgamated with Columbia Graphophone Company to form EMI, which took over the studios. The studios were then known as EMI Studios until EMI formally changed their name to Abbey Road Studios in 1970.

Studio Two at Abbey Road became a centre of rock music in 1958 when Cliff Richard and the Drifters (later Cliff Richard and The Shadows) recorded "Move It" there, arguably the first European rock and roll single.

The zebra crossing in 2007Abbey Road Studios is most closely associated with the Beatles, who recorded almost all of their albums and singles there between 1962 and 1970. The Beatles named their 1969 album, Abbey Road, after the street where the studio is located (the recording studio would only be named Abbey Road after the Beatles record in 1970). The cover photo for that album was taken by Iain Macmillan outside Abbey Road Studios, with the result that the pedestrian zebra crossing outside the studio, still located in exactly the same spot today, has become a place of pilgrimage for Beatles fans from all over the world. It has been a long-standing tradition for visitors to pay homage to the band by writing on the studio fence in front, although it is painted over monthly. In December 2010 the zebra crossing at Abbey Road was given a Grade II listed status.

Pink Floyd recorded most of their late 1960s to mid-1970s albums, returning only in 1988 for mixing and overdubbing subsequent albums.

The Shadows named their Live At Abbey Road album after the studio, with the cover spoofing The Beatles' album.

Notable producers and sound engineers who have worked at Abbey Road include Sir George Martin, Geoff Emerick, Norman "Hurricane" Smith, Ken Scott, Mike Stone, Alan Parsons, Peter Vince, Malcolm Addey, Peter Bown, Richard Langham, Phil McDonald, John Kurlander, Richard Lush and Ken Townsend, who invented the groundbreaking studio effect known as automatic double tracking (ADT). The chief mastering engineer at Abbey Road was Chris "Vinyl" Blair, who started his career early on as a tape deck operator. Blair died on 7 November 2005.