Background
She was the fourth daughter of James Cunningham, 7th Earl of Glencairn and Margaret, daughter of Sir Colin Campbell of Glenurquhy, a family noted for their early commitment to Protestantism. Her sister was the memoirist Lady Margaret Cunningham.
She married James Hamilton, 2nd Marquess of Hamilton on 30 January 1603. They had two sons, James Hamilton, 1st Duke of Hamilton (1606–1649) and William Hamilton, 2nd Duke of Hamilton (1616–1651) and three daughters, Lady Anne Hamilton (married Hugh, 7th Earl of Eglinton), Lady Margaret Hamilton (married John, Earl of Crawford and Lindsay) and Lady Mary Hamilton (d 1633) (married James Douglas, 2nd Earl of Queensberry).