The recent renovation of a house in Chajul in western Guatemala has revealed an unparalleled set of wall paintings, most probably from the Colonial period (AD 1524–1821). The iconography of the murals combines pre-Columbian elements with imported European components in a domestic rather than a religious setting, making them a unique example of Colonial-period art. Here, the authors present the results of iconographic, chemical and radiocarbon analyses of the Chajul house paintings. Dating to the seventeenth to eighteenth centuries AD, the paintings may be connected to a revival of the local religious organisation (cofradías) in the context of waning Spanish colonial control.