The third earl was a minor at his father's death, and became a ward of Cardinal Wolsey. He took his seat in the House of Lords in the parliament that met on 3 Nov. 1529, and on 13 July 1530 he was one of the peers who signed the letter to the pope petitioning him to grant Henry VIII's divorce. In 1532 he was present with Henry at his interview with Francis I at Boulogne. He was made a knight of the Bath on 30 May 1533, and on 1 June following he officiated as cupbearer at the coronation of Anne Boleyn. He took a prominent part in suppressing the northern rebellions in 1536 and 1537 (Letters and Papers of Henry VIII, ed. Gairdner, vols. xi. and xii. passim). In 1542 he accompanied Thomas Howard, third duke of Norfolk, on his raid into Scotland. He was elected K.G. on 17 Feb. 1546–7, and three days later bore the sword ‘curtana’ at the coronation of Edward VI. He was, however, strongly opposed to religious change, and protested in the House of Lords against the bills confirming the new liturgy (10 Dec. 1548), for the destruction of the old service books (December 1549), compelling attendance at divine service (January 1552–3), and legalising the marriage of priests (March 1552–3). In June 1551 it was reported that he had been commanded to ‘renounce his title of the Isle of Man,’ but refused, and was preparing to resist by force (Cal. State Papers, For. i. 119–20). Nevertheless, he was on 9 Aug. 1551 sworn a privy councillor on condition of attending only when specially summoned, and in the same year he was one of the parties to the peace with Scotland. He took little part in the proceedings of the council, but in December 1551 he was one of the peers who tried Somerset, while his eldest son was one of the principal witnesses against the duke. On 16 May 1552 he was appointed lord lieutenant of Lancashire.
Derby naturally welcomed the accession of Queen Mary, and was one of her earliest adherents. On 17 Aug. 1553 he was made a regular member of the privy council, which he frequently attended, and in the same month was placed on a commission to investigate Bonner's deprivation of the bishopric of London. He was created lord high steward for the coronation of Mary on 1 Oct. and bore the sword ‘curtana’ at that ceremony. On 11 Nov. following he was made a special commissioner for the trial of Lady Jane Grey and others, and during Mary's reign he frequently took part in the proceedings against heretics, John Bradford (1510?–1555) [q. v.] being one of the victims of his activity (Foxe, Actes and Mon. vol. vii. passim; Maitland, Essays on the Reformation). He attended Philip of Spain at his landing on 19 July 1554, and on 30 May 1557 he was appointed captain of the vanguard to serve against the Scots. He was one of those summoned to attend Queen Elizabeth on her entry into London in November 1558, and before the end of the year became a member of Gray's Inn. He was retained as a member of the privy council, was appointed chamberlain of Chester on 16 April 1559, visitor of the churches in the province of York on 24 June 1559, commissioner for ecclesiastical causes in the diocese of Chester on 20 July 1562, and lord lieutenant of Cheshire and Lancashire on 18 Nov. 1569. But though he often took part in proceedings against recusants and gave the government timely warning of the insurrection of 1569, his sympathies and connections rendered him an object of suspicion to Elizabeth. The queen's enemies counted on his support (cf. Cal. State Papers, Dom. Addenda, 1566–79, pp. 371–2), and his sons, Edward and Thomas, were in 1571 implicated in an attempt to release Mary Queen of Scots from Tutbury (Hatfield MSS. i. 505–76). Derby died at Lathom House on 24 Oct. 1572; he had been noted for his splendid hospitality, and his funeral at Ormskirk on 4 Dec. 1572 was one of the most magnificent on record (cf. The Derby Household Books, Chetham Soc.; Cal. State Papers, Dom. 1547–81, p. 455; Collins, Peerage, iii. 55–62). His will, dated 24 Aug., was proved on 21 Nov. 1572. An engraving of an anonymous portrait of Derby belonging to the present Earl Derby is given by Doyle.
Derby was thrice married. His first wife was Katherine (her name is given in the peerages as Dorothy), daughter of Thomas Howard I, second duke of Norfolk, who on 21 Feb. 1529–30 received a pardon ‘for the abduction of Edward, earl of Derby, and marriage of the said Edward to Katherine, daughter of the said Thomas, without royal license’ (Letters and Papers, iv. 6248, art. 21). By her Derby had issue Henry Stanley, fourth earl [q. v.], Sir Thomas Stanley (d. 1576), and Sir Edward (d. 1609); and four daughters. His second wife was Margaret, daughter of Ellis Barlow of Barlow, Essex, by whom he had one son and two daughters. She died on 23 Feb. 1558–9, and an epilogue on her death, by Richard Sheale, is printed in the ‘British Bibliographer,’ vol. iv. (cf. Stanley Papers, i. 14). His third wife was Mary, daughter of Sir George Cotton of Combermere Abbey, Cheshire, who afterwards married Henry Grey, earl of Kent, and died without issue on 16 Nov. 1580.