In Malaysia, the number of registered organ donors are relatively small in comparison to other countries. This study aimed to investigate the awareness, knowledge and practices on organ donation among public in Sarawak. A cross-sectional study was conducted in urban and rural parts of Bintulu, Sarawak. Questionnaires consisting of socio-demography (5 items), awareness (4 items), knowledge (6 items) and practice (5 items) questions were distributed using random and snowball sampling techniques. A total of 327 respondents (age = 33.27 ±10.06 years; urban area = 50.2%; male = 59.0%; Christian = 48.9%; Bumiputra Sarawak = 53.2%) were enrolled. Urban respondents have demonstrated better knowledge (definition = 24.1% vs 23.6%; treatment = 42.2% vs 41.6%; side effects = 45.2% vs 32.9%; from living donor = 80.1% vs 70.8%; from dead donor = 71.7% vs 53.4%; organs can be donated = 19.9% vs 27.3%) compared to rural respondents. Urban respondents possessed better awareness (awareness = 86.1% vs 73.9%; source of information = 39.2% vs 31.7%; need = 70.5% vs 50.3%) than their counterparts. Subsequently, urban respondents reported better practice (agree = 68.7% vs 60.2%; registered = 2.4% vs 1.2%; donate after death = 27.7% vs 25.5%; without permission = 20.5% vs 18.0%; for loved ones = 72.9% vs 64.6%) than rural respondents. Conclusion: Population in urban area are more aware, have higher knowledge and better practice regarding organ donation. Health education on organ donation should be encouraged specifically among rural residents to enhance the number of registered organ donors in Malaysia.