Nowadays exist has many studies dealing with employee motivation. The aim of our work was to find out whether there are statistically significant differences in employee motivation depending on gender and age. Considering the amount of data collected through a questionnaire of 30,772 respondents in the years since 2012 to 2017, we worked from the point of view of the respondents with the top five most important motivated factors - basic salary, good working collective, fair assessment, job security and workplace atmosphere. From the used statistical methods, we have subsequently confirmed the alternative hypothesis, that between the independent variables age and gender and the level of motivation induced by the individual motivational factors is statistically significant depend. We compared after that the results of our results with the results of various authors in the part of called discussion.