Motivational factors play an important role in increasing employee job satisfaction. Satisfied employees in return can help in improving organizational performance. The objective of following research is to analyze the effects of motivational factors on job satisfaction of employees. This is an exploratory study based on primary data. The primary data has been collected from non-academic staff of University of the Punjab, which is one of the biggest universities of Pakistan. The study of Herzberg et al. (1959) has been widely validated and well renowned among research community. This theory tests hygiene and motivator factors and impact of personal and job characteristics on work perceptions and job satisfaction. Structural equation modeling technique has been applied to test hypothesis, SPSS 16.0 has also been adopted for basic analysis purposes. The results are being questionable to verify Herzberg's theory. The study concludes that intrinsic motivational factors are having significant relationship with employee job satisfaction, whereas hygiene International Journal of Business and Management March, 2010 71 (extrinsic) factors are not having any significant relationship with employee job satisfaction. Moreover, significant difference was observed between gender, qualification, experience, job characteristics and job satisfaction. This study offers useful information as it provides both, the practical implications for professionals working on authoritative posts within Public Sector University set up in Pakistan, and theoretical implications for researchers interested in exploring job satisfaction in a higher education context.