Son of William Barlow and Agatha Wellesbourne, he was born at St David's when his father was bishop of that diocese, and was educated at Balliol College, Oxford.
About 1573 he entered into holy orders, and was made a prebendary of Winchester (1581) and rector of Easton.
In 1588, while maintaining his position at Winchester, Barlow began an association with Lichfield as prebend of Colwich, which in the following year was transferred to the 'golden prebend' of Sawley with which came the position of treasurer of Lichfield Cathedral.
He afterwards became a chaplain to Henry Frederick, Prince of Wales, son of James I, and finally archdeacon of Salisbury (1615).
Barlow died 25 May 1625, and was buried in the chancel of his church at Easton.