This study conceptualizes e-service quality with respect to a hotel website and develops a corresponding measurement scale. Using a mixed methodology research design, 29 semi-structured interviews first defined customers’ perceptions of hotel websites and identified the important areas for investigation. A subsequent online survey of people who had recently visited a hotel website resulted in 843 usable responses, from which the e-service quality measurement scale was developed. The measurement scale for hotel websites comprised 24 items across six factors: functionality, atmospheric quality, reliable information, locality information, customer reviews, and emotional engagement. The investigation found two higher order factors (environment and process quality) to measure e-service quality for hotel websites. The newly developed scale gives a better understanding of e-service quality in the hotel context and enables hotel practitioners to improve hotel website quality.