The aim of this study was to investigate the reliability and validity of the Rejection sensitivity Questionnaire (RSQ) in university students. The research sample consisted of 325 undergraduate students from the University of Isfahan (170 female, 150 male). Who were selected randomly through cluster sampling. The instruments used in this research were Rejection sensitivity Questionnaire (RSQ), Penn State Worry Questionnaire, and Rosenberg Self-Esteem Questionnaire. In order to assess the questionnaire's reliability, Cronbach's alpha was used. Correlation coefficient, exploratory and confirmatory factor analysis were used to assess construct validity of RSQ. Cronbach’s Alpha of the questionnaire was 0.84. In exploratory factor analysis, 2 factors (1- response expectation, 2- worry of unacceptance of request) were extracted which explained 28.28% of total variance. Confirmatory factor analysis confirmed these two factors. Penn State Worry Questionnaire and Rosenberg Self-Esteem Questionnaire were used to assess the convergent and divergent validity of the RSQ respectively. Both of them correlated significantly with RSQ. There was no difference between RSQ scores in males and females. The results of this study showed that the RSQ had satisfactory reliability and validity coefficients and can be a suitable tool for measuring Rejection sensitivity in Iranian students in future research.