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Barnardiston as representatives of Suffolk, and an attempt was made on the part of the royalists to discredit the importance of the election by the assertion that ‘they would not have been chosen if there had been any gentlemen of note, for neither Ipswich had any great affection for them nor most of the country; but there were not ten gentlemen at this election’ (Cal. Dom. State Papers, 4 March 1627–8). During the long interval between the parliament of 1629 and the summoning of the short parliament in 1640, Sir Nathaniel seems to have lived quietly at Ketton. He had married Jane, daughter of Sir Stephen Soame, knight, and alderman of London, who was lord mayor in 1597–8, and had by her a large family, in whose religious education he was deeply interested. His piety at home (he prayed thrice a day), and his benevolence to ministers of religion, gave him a wide reputation among the puritans of the eastern counties. ‘He had ten or more servants so eminent for piety and sincerity that never was the like seen all at once in any family.’ He encouraged in his parish catechetical instruction in religion; and he attended with his children the religious classes held by Samuel Fairclough, the rector of Ketton; replied himself to the questions that his sons and daughters were unable to answer, and urged his neighbours, both rich and poor, to follow his example. In 1637 his wife, Lady Barnardiston, gave 200l. ‘to be bestowed by his direction’ to Mr. Marshall, vicar of Finchingfield, who was described by the vicar-general of London as governing ‘the consciences of all the rich puritans in these parts and in many places far remote’ (Cal. Dom. State Papers, March 1636–7). On 14 April 1640 Sir Nathaniel was returned to the Short parliament for his county, and in October he was elected to the Long parliament for the same constituency (cf. Harl. MS. 165, No. 5). In 1643 he took the covenant, became a parliamentary assessor for Suffolk, and joined the Eastern Counties' Association. He does not appear to have taken any active part in the war, but he was in close relations with the leaders of the parliament (, Memorials, i. 467). He subscribed 700l. and lent 500l. to the parliament for the reduction of the Irish rebels; the latter sum was ‘to be repaid with interest at the rate of eight per cent.’ out of the first payments of the parliamentary subsidy of 400,000l. levied in 1642. On 10 May 1645 he petitioned parliament to repay the greater part of his loan, for which he declared he had special occasion, and his request was formally granted (Commons' Journal, iv. 133; Lords' Calendar in Hist. MSS. Com. Rep. vi. 59 a). Shortly after the execution of the king, Sir Nathaniel's health broke down, and he retired to Ketton to prepare for death. He devoted himself unceasingly to religious exercises during his last two years (1651–1653), and read constantly Baxter's ‘Saint's Everlasting Rest.’ About 1652 he removed to London for the convenience of his doctors, and died at Hackney on 25 July 1653. ‘His corpse being carried down from London was met about twenty miles from his own house by 2,000 persons, most of them of quality; and his funeral at Ketton on 26 Aug. Following was attended by many thousands.’ The sermon was preached by Samuel Fairclough, the rector, his intimate friend and adviser, who had been presented to the living 26 Jan. 1629–30, and it was published under the title of ‘Hagioi Axioi or the Saints Worthinesse and the Worlds Worthlesnesse, both opened and declared in a Sermon preached at the Funerall of that eminently religious and highly honoured Knight, Sir Nathaniel Barnardiston,’ with a dedication to Lady Jane Barnardiston and her children. The sermon, which is a full memoir of the life of Sir Nathaniel, was reprinted in Samuel Clark's ‘Lives of Sundry Eminent Persons in this Later Age’ (1683). A collection of elegies on his death was issued, later in 1653, under the title of ‘Suffolks Tears, or Elegies on that renowned knight, Sir Nathaniel Barnardiston. A Gentleman eminent for Piety to God, love to the Church, fidelity to his Country.’ Twenty-two English poems, twelve Latin, and one Greek are included, which are all of very mediocre quality. One of the best is ‘The Offering of an Infant Muse’ (p. 39), signed ‘Nath. Owen, anno ætat. 12°.’

Lady Jane Barnardiston, who shared her husband's religious fervour, was buried at Ketton, 15 Sept. 1669. Of Sir Nathaniel's eight sons, the eldest, Sir Thomas, and the third, Sir Samuel, both attained political eminence [see, and ]. Another of his sons, John, has been identified with the Mr. Barnardiston, member of the committee of parliament in the eastern counties, who was seized by the royalists at Chelmsford in 1648; was imprisoned in Colchester Castle at the time that the parliamentarians were besieging it; was released in order to negotiate terms with Sir Thomas Fairfax; and finally signed articles (20 Aug. 1648) which assented to the execution of two royalist leaders, Sir George Lisle and Sir Charles Lucas