The motives behind self-employment and the differences between women and men are a hot topic in entrepreneurship debate. This paper empirically explores the interaction of three types of motivation on the entrepreneurial activities of women and men in 24 European countries: opportunity-driven motivation, necessity-driven motivation and mixed motivation. Employing a dynamic method, a panel data analysis in the timeframe from 2009 to 2012 is conducted. In this regard, the paper explores entrepreneurship at the two levels of established businesses (EBs) and total early-stage entrepreneurial activities (TEAs). The findings suggest that all three motivational dimensions positively influence females’ self-employment at both levels. Also, the findings reveal that, at the established business level, there is a significant and positive relationship between entrepreneurship by men and opportunity-seeking motivation.