Empirically based tools to map education for sustainable development within school organisations are not readily available, which is both a cause and a consequence of the scarce empirical and quantitative research on school organisations and education for sustainable development. In present study, the Education for Sustainable Development School Organisation Questionnaire (ESD-SOQ) was developed and validated. This instrument measures the organisational characteristics of an ESD-effective school. During the development and validation of the ESD-SOQ different steps were taken. First, data from prior research was consulted to draft the initial scales. These were further refined via cognitive interviews (n = 3) and consulting critical peers (n = 7). Next a pilot study (n = 108) was set out to retain the best items per scale and thus reduce the size and load of the questionnaire. Finally, a large-scale validation study (n = 764) ensured the reliability, the content validity, the cognitive validity and the construct validity of the final instrument. Based on the findings of our development and validation study, ESD-SOQ proves to be a valid and reliable instrument for the investigation school organisational characteristics related to education for sustainable development.