Acceptability (of healthcare services) is an important construct that lacks a consistent definition within research. Addressing this issue, a systematic review led to the Theoretical Framework of Acceptability. In this study, we describe the development (based on the Theoretical Framework of Acceptability) and validation of the Digital Health Acceptability Questionnaire. Nineteen items aligning with the Theoretical Framework of Acceptability were developed. Two versions of the questionnaire measuring telehealth acceptability by telephone (N = 644) and videoconference appointment (N = 425), were administered to a nationally representative survey of consumers in Australia. Two exploratory factor analyses (Oblimin rotation) were conducted for each scale (telephone/videoconference). Two-factor solutions (5 items each) were found for both (telephone/videoconference) acceptability questionnaires: (a) attitude toward the service as a means to address healthcare needs and affective attitude and (b) individual capacity and effort to use telehealth. Before rotation, Factor 1 of the telephone scale (α = 0.92) measured 56.18% of the variance and Factor 2 (α = 0.86) measured 14.17%. Factor 1 of the videoconference scale (α = 0.90) measured 56.68% of the variance and Factor 2 (α = 0.85) measured 10.63%. The full10-item acceptability questionnaire showed excellent internal consistency (telephone: α = 0.91 and videoconference: α = 0.92). The 2-dimensional Digital Health Acceptability Questionnaire is a brief survey based on research evidence and validated in a large Australian sample.