There is a traditional story (now perhaps discredited as a fictionalised account by Sir Walter Scott[7]) that James was an especial enemy of the Regent's party because his wife and child were evicted from Woodhouselee.

The ghosts of his tragic wife and child are said to still haunt the site of the old castle and her frenzied and terrifying screams have been heard by those dwelling nearby.

[10] However, the main motive for the assassination was rooted in the political rivalries in Scotland at the time, and James Hamilton's uncle, John Hamilton, Archbishop of St. Andrews, at least had prior knowledge of the plot[11] (It has been suggested that the Archbishop and James's mother Christian Shaw may have been born to the same mother, or that "uncle" was merely a courtesy title).

Shortly after Moray's assassination, his secretary Master John Wood was also murdered, which second killing has been taken by commentators since the contemporary George Buchanan to support the interpretation that this was a political crime and not a personal feud.

[12] James Hamilton decided to assassinate Moray and travelled to the Borders, Edinburgh, York, London, Perth, Glasgow and Stirling without an opportunity arising.

Knox's source had told him that the attempt would be on the High Street, near the cross, and an alternative route to avoid the Archbishop's house was suggested in vain.

[15] In Ritchie's story, James placed feathers on the ground to deaden his footsteps, hanging a black cloth on the wall to hide his shadow and obtaining a brass match-lock carbine with a rifled barrel for accuracy.

[19] After a desperate ride, closely pursued by the Regent's men, James made it to the safety of his triumphant kin in Hamilton.

Regent Moray dismounted, wounded below his navel, walked to his lodging and died the same day, according to the Diurnal of Occurrents at Linlithgow Palace in the hour before midnight.

[23]The 17th-century historian David Calderwood described the shot in more detail; "with a hacquebut, through a tirleis window (shuttered), from a stair whereupon were hunge sheets to drie, but in truthe, to hide the smooke, and make the place the lesse suspected".

According to David Calderwood, the next night after Moray's death, Walter Scott of Buccleuch and Thomas Ker of Ferniehirst raided the English border deliberately to help the conspiracy.

[27] Moray was buried at St Giles Kirk in Edinburgh, his body carried by six earls and lords and his standard of the Red Lion of Scotland by William Kirkcaldy of Grange.

John Knox had prohibited funeral sermons on the grounds that they glorified the deceased and displayed distinctions between rich and poor, however the ban was lifted on this occasion.

[30] One of the finest remaining brasses in Scotland commemorates the murdered Earl of Moray, and is located in Saint Giles Kirk, Edinburgh.

He was asked to assassinate Gaspard II de Coligny; however, he refused, stating that a man of honour was entitled to settle his own quarrels, but not to murder for others.

One letter asked a servant of Mary Queen of Scots to send him financial aid because he had lost "all he had to live on for her Majesty's service".

[36] The main perpetrator, James Hamilton of Bothwellhaugh, was still in France: in the letters of the English ambassador Lord Cobham his title was spelled "Bodilaugh", perhaps an indication of how the name was said.

[37] Christian Shaw, James's mother, the widow of David Hamilton of Bothwellhaugh, was charged with assisting the assassination in February 1571 at Lanark.

On 16 June 1582, George Hume of Spott was charged with, among other crimes, helping the assassination by welcoming the "actual committaris", named as James Hamilton and his brother John, the Provost of Bothwell, after the shooting, at his house at Neidpath-Head.

The Regent's widow Agnes Keith and daughter Elizabeth Stuart were represented at the proceedings, and Spott was acquitted of the other charges,[38] James Hamilton's brother David, his wife Isobel Sinclair and her sister, Alison Sinclair, David's wife, were fully restored to the forfeited inheritance of Woodhouselee in January 1592, apparently despite the resistance of the Lord Clerk Register.