This paper aims to define the motivational preferences of employees at large concerns in the Czech Republic. The paper presents the results of empirical research at the level of employee motivation. The research was conducted at the end of 2018. Four hundred and forty-four companies were addressed. A questionnaire was distributed to companies in the manufacturing industry. The methods of data processing used were statistical mathematical methods, specifically the Chi-squared test and factor analysis. The study demonstrates that the possibility of career education and the transparency of remuneration have a large effect on the motivation of employees in the Czech environment; these results are subsequently compared with extensive theoretical research. The findings made by this study may be used as a tool to increase employee motivation, which has a direct influence on both employee performance and the performance of the company as a whole.