A gendered analysis of Roman inquisitorial tribunals reveals how underlying expectations of masculine and feminine attitudes and behaviours helped shape the practices of inquisitors, defendants, accusers and witnesses, both in Rome and in the various peripheral tribunals of the Roman Inquisition. This gender-focused study reinforces some current approaches and arguments that peripheral tribunals and their local inquisitors had leeway for procedural developments away from central Roman dictates. Tribunals also had to make their own decisions on how to react to defence ploys, especially women’s defence strategies, not anticipated by central manual writers. This study pays close attention to how inquisitors, accusers and witnesses described heretical bodies. It also argues that both urban spaces and speech in early modern cities were gendered, a fact that had important implications for how women were accused and tried, and how they attempted to defend themselves, in tribunals of the Roman Inquisition.