Research Background: Occupational stress results from various interactions of the worker and the environment of the work they perform their duties. Occupational stress results from the interaction of the worker and the conditions of work. Occupational stress is stress involving work. In this study, an investigator has focused on finding out the impact of region and gender differences on occupation stress of employees. Objectives: The study objective was, to find out the impact of region and gender differences on occupation stress of employees. Other, objective was, to study the significant difference in level of occupational stress among the male and female urban as well as rural employees. Procedure: The study one hundred twenty samples were selected from the clerical based employees in the State of Maharashtra. Out of this sample, sixty clerical based employees were selected from urban region and same way sixty clerical based employees were selected from rural region. The sample age range was between 35 and 60 years. The purposive sampling technique was used for the selection of the samples. The study type of region and type of gender were independent variable and level of occupational stress was dependent variable. The study Occupational Stress Index developed by Dr. A. K. Srivastava and Dr. A. P. Singh has been used. Conclusions: It is concluded that, insignificant difference found in occupational stress among the urban and rural employees. The female employees have found high level occupational stress than male employees. The urban female employees found have high level occupational stress than urban male employees, other, hand side the rural female employees found have high level occupational stress than rural male employees. Application: The present study findings can be beneficial and helpful for counselors, society member, psychologists to understanding impact of region and gender differences on occupational stress of individuals. Also, these study findings can be beneficial for national and international research scholars.