This article examines the Domine, Quo Vadis? painting by Annibale Carracci (1560–1609) that hangs in the National Gallery, London. Painted between 1601–1602, the panel depicts the apocryphal encounter between Christ and St Peter on the Appian Way, as the latter flees from persecution by Nero’s guards in Rome. Peter asks Christ, ‘Master, where are you going?’, to which Christ responds, ‘I am going to Rome, to be crucified anew’. Following the apparition, Peter turns back and sacrifices himself instead. Carracci diverges significantly from the traditional artistic renderings of the incident, both in terms of narrative strategy and scenic composition. It will be argued that his treatment of the subject can be interpreted as a direct address to the patron of this work, Cardinal Pietro Aldobrandini (1571–1621). Playing upon the moral underpinning of the subject, Carracci devises an innovative composition that is tailored to stimulate personal introspection in the Cardinal. However, the piece performs the equally important function of projecting outwards the powerful and learned status of its owner. Carracci’s work flatteringly implies an erudite patron through its overt references to Raphael, Michelangelo and the ancient past of Rome. The approach of Raphael in particular to classical antiquity was admired by Cardinal Aldobrandini and his personal secretary, Giovanni Battista Agucchi (1570–1632). The idealising classicism of his late career earned Carracci the praise of Agucchi, whose role in shaping the taste of the Cardinal must be acknowledged. The Domine, Quo Vadis? fulfils Agucchi’s desire that artists should have an idea of things ‘not as they are, but as they ought to be’. Carracci also inserts a fictive paesaggio classico into his Domine, Quo Vadis?. This emerging Seicento pictorial type, to which Cardinal Aldobrandini was especially partial, is the final confirmation of a work destined unmistakably for a unique audience.