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The current study examined several aspects of validity evidence for the Faculty Readiness to Teaching Online scale. Data were collected from 307 faculty in higher education. Construct validity was evidenced through an exploratory factor analysis (EFA) and a reliability coefficient. Differential validity was explored using the differential item functioning (DIF) analysis to explore potential bias in items due to gender. In addition, convergent validity was tested by correlating FRTO with years of teaching online, the primary modality of instruction, and whether there is required training in online teaching by the institution. The findings from validation showed that new factors emerged and also demonstrated differences based on instructors gender, as well as based on teaching experience and primary modality of teaching.