Dowland worked for many years at the court of Christian IV of Denmark. He returned to England in 1606 and in 1612 secured a post as one of James I's lutenists. Most of Dowland's music is for his own instrument, the lute. It includes several books of solo lute works, lute songs (for one voice and lute), part-songs with lute accompaniment, and several pieces for viol consort with lute. The poet Richard Barnfield wrote that Dowland's "heavenly touch upon the lute doth ravish human sense."

One of his better known works is the lute song "Flow My Tears". He later wrote what is probably his best known instrumental work, Lachrimae or Seaven Teares Figured in Seaven Passionate Pavans, a set of seven for five viols and lute, each based on "Flow My Tears." It became one of the best known pieces of consort music in his own time. His pavane "Lachrymae antiquae" was also one of the big hits of the seventeenth century.

Dowland's music often displays the melancholia that was so fashionable in music at that time. He wrote a consort piece with the punning title Semper Dowland, semper dolens (always Dowland, always doleful), which may be said to sum up much of his work.

Dowland's song, Come Heavy Sleepe, the Image of True Death, was the inspiration for Benjamin Britten's Nocturnal after John Dowland for guitar, written in 1964 for the guitarist Julian Bream. This work consists of eight variations, all based on musical themes drawn from the song or its lute accompaniment, finally resolving into a guitar setting of the song itself.