James Hamilton of Bothwellhaugh and Woodhouselee was a Scottish supporter of Mary, Queen of Scots, who assassinated James Stewart, 1st Earl of Moray, Regent of Scotland, in January 1570. He shot Moray from the steps of his uncle Archbishop John Hamilton's house in Linlithgow.

James Stewart, 1st Earl of Moray was a member of the House of Stewart as the illegitimate son of King James V of Scotland. At times a supporter of his half-sister Mary, Queen of Scots, he was the regent of Scotland for his half-nephew, the infant King James VI, from 1567 until his assassination in 1570. He was the first head of government to be assassinated with a firearm.