John Humphrey Barbour was born on May 29, 1854, the son of Henry S. Barbour (1822-1891), town clerk in Torrington, Connecticut, and his wife, Pamela J. Bartholomew (1827-1899). John Barbour had one sister, Lucy Amelia (b. 1863), and one brother, Edward Willis (1857-1861). John studied at Amherst College in Amherst, Massachusetts, before graduating from Trinity College, Dublin, in 1873. In 1876, he earned a degree in divinity from Berkeley Divinity School in New Haven, Connecticut, and he began his ministerial career at the Episcopal Church in Parkville, Connecticut. He became a professor at Berkeley Divinity School around 1889 and also served as the school's librarian. He married Annie Gray on May 7, 1878, and they had three surviving children: Ellen Gray, Henry Gray, and Paul Humphrey. Paul also became a minister. John Humphrey Barbour died on April 29, 1900.