While female diplomatic representation was almost non-existent during the socialism in the Czech Republic, a number of women diplomats has been slowly increasing since the nineties. Women are currently solidly represented in the Ministry of Foreign Affairs but only a small fraction serves in diplomatic missions and reaches higher diplomatic positions. This paper tests the findings of recent gender research studies in international relations which explore the statutes and challenges encountered by female diplomats. Based on the current state of the research, this paper works with two hypotheses. Firstly, only a small percentage of female diplomats, approximately 15%, reach the status of ambassadors and they are positioned in less prestigious missions. Secondly, even though prejudice towards abilities of female diplomats still exists, it is mainly due to practical problems of balancing family and professional life that the women are not able to participate and to reach higher diplomatic positions in a greater rate. The presented analysis works with both quantitative and qualitative data and draws on theories of gender and positional status. It explores official statistics and documents of the Czech Ministry of Foreign Affairs (MFA) as well as existing legal regulations on the public and foreign service with the focus on the latest reforms which added new positive measures. Qualitative part of the research is based on semi-structured interviews with the Czech female and male ambassadors, employees of the MFA human resources department, the MFA gender coordinator and the higher MFA representatives. Final results of the research fully confirm stated hypotheses and add learnings on specificities of the Czech context of the diplomatic work.