Originally from Devon, where he was born into a farming family, Lane moved to London already in his teens. While working as a clerk at the Railway Clearing House, he acquired knowledge as an autodidact.
After entering the London book trade he became co-founder of The Bodley Head, originally a firm that dealt with antiquarian books. They later went into publishing. Lane is mainly associated with publishing controversial and audacious texts, especially for a small, sophisticated audience. Examples are the periodical The Yellow Book (1894 - 1897) and Lane's Keynote Series, which included contentious material such as Grant Allen's novel The Woman Who Did (1895), Victoria Crosse's immediate reaction to it, the novel The Woman Who Didn't (1895), and H.G. Wells's novel about his affair with Amber Reeves, The New Machiavelli (1911).
On 13 August 1898, John Lane married Annie Philippine King, the widow of Tyler Batcheller King and the daughter of Julius Eichberg. Annie Lane was author of To Thee, O Country (national hymn) and of the books Brown's Retreat, Kitwyk (published by John Lane in 1903), The Champagne Standard, Talk of the Town and According to Maria. John Lane died of pneumonia on 2 February 1925 at his London home, 8 Lancaster Gate Terrace, Bayswater, London. He was cremated at Golders Green, and his ashes were interred at St Nectan's Church, Hartland, Devon. His nephew, Allen Lane, founded Penguin Books.
In the churchyard of St Nectan's, Stoke, near Hartland in Devon, there is a stone seat commemorating various members of the Lane family.