Isaac Ambrose (1604 - January 20, 1663/1664) was an English Puritan divine, the son of Richard Ambrose, vicar of Ormskirk, and was probably descended from the Ambroses of Lowick in Furness, a well-known Roman Catholic family.

He entered Brasenose College, Oxford, in 1621, in his seventeenth year. Having graduated B.A. in 1624 and been ordained, he received in 1627 the little cure of Castleton in Derbyshire. By the influence of William Russell, earl of Bedford, he was appointed one of the king's itinerant preachers in Lancashire, and after living for a time in Garstang, he was selected by the Lady Margaret Hoghton as vicar of Preston. He associated himself with Presbyterianism, and was on the celebrated committee for the ejection of "scandalous and ignorant ministers and schoolmasters" during the Commonwealth.

So long as Ambrose continued at Preston he was favoured with the warm friendship of the Hoghton family, their ancestral woods and the tower near Blackburn affording him sequestered places for those devout meditations and "experiences" that give such a charm to his diary, portions of which are quoted in his Prima Media and Ultima (1650, 1659). The immense auditory of his sermon (Redeeming the Time) at the funeral of Lady Hoghton was long a living tradition all over the county. On account of the feeling engendered by the civil war Ambrose left his great church of Preston in 1654, and became minister of Garstang, whence, however, in 1662 he was ejected with the two thousand ministers who refused to conform. His after years were passed among old friends and in quiet meditation at Preston. He died of apoplexy about 20 January 1663/4.

As a religious writer Ambrose has a vividness and freshness of imagination possessed by scarcely any of the Puritan Nonconformists. Many who have no love for Puritan doctrine, nor sympathy with Puritan experience, have appreciated the pathos and beauty of his writings, and his Looking to Jesus long held its own in popular appreciation with the writings of John Bunyan.

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• This article incorporates text from the Encyclopædia Britannica, Eleventh Edition, a publication now in the public domain.