Extensive job stress and workload are main causes of frustration and reduced efficiency among workers and despite negative effects of these factors on employees’ performance, related studies in rehabilitation staff are limited. The aim of this study was to evaluate job stress, workload and their related factors in rehabilitation staff. In this cross-sectional study, 150 employees in three rehabilitation professions including physical therapy (PT), occupational therapy (OT) and orthotics and prosthetics (OP) were selected randomly out of all the professionals in these careers in Tehran in 2013. Socio-demographic questionnaire, job stress inventory (JSI) and the NASA task load index (NASA-TLX) were applied as research tools. To analyze the data, descriptive and inferential statistics were applied using SPSS.19 software.The results indicated that job stress in PT and OP is high and although the difference in the mean job stress score between PT and OP was not significant, there is a significant difference between job stress in these groups and OT. The amount of workload in PT was significantly higher than OP and OT, but the difference in workload between OP and OT was not significant. Job experience had positive correlation with workload and job stress. No significant differences were identified in mean scores of job stress and workload with regard to gender, marital status and level of education. Significant higher job stress was found in public sector employees compared to private sector ones and in temporary employees compared to those who had permanent contract. Due to high levels of perceived job stress and workload in rehabilitation staff, recognizing effective factors for modifying these problems and assessment of work related disorders among these professionals shouldbe given a priority.