Isotachophoresis (ITP) utilizes the formation of a permanently sharp interface between electrolytes containing substances of different mobility. Due to its specific properties, especially its ability to concentrate analytes from very diluted samples, this technique found its firm place in the field of separation methods, maintaining it successfully for over 50 years. We can argue with certainty that Czech and Slovak scientists contributed significantly to the development of ITP. In this short review, we have tried to describe the historical development of ITP in Czechoslovakia, the Czech Republic, and Slovakia. The text is arranged more or less chronologically and structured according to the individual laboratories and teams.