Isaac Ambrose (1604 – 20 January 1664) was an English Puritan divine. He graduated with a BA from Brasenose College, Oxford, on 1624. He obtained the curacy of St Edmund’s Church, Castleton, Derbyshire, in 1627. He was one of king's four preachers in Lancashire in 1631. He was twice imprisoned by commissioners of array. He worked for the establishment of Presbyterianism; successively at Leeds, Preston, and Garstang, whence he was ejected for nonconformity in 1662. He also published religious works.