Cieľom práce bola izolácia Na+/K+-ATPázy a optimalizácia merania kinetiky Na+/K+-ATPázy metódou stopped-flow. Teoretická časť práce sa venuje membránovému transportu, štruktúre a mechanizmu fungovania Na+/K+-ATPázy. Ďalej sú rozoberané jednotlivé metódy, ktoré sa využívajú na sledovanie kinetiky ATPáz. V praktickej časti je popísaná izolácia Na+/K+-ATPázy a meranie kinetiky Na+/K+-ATPázy. Pre meranie bolo použité fluorescenčné farbivo RH421. Bola sledovaná zmena intenzity fluorescencie po rýchlom zmiešaní dvoch roztokov, kde jeden obsahoval farbivo inkubované s Na+/K+-ATPázou a druhý len ATP. Pri optimalizácii metódy sa vyskytol problém s rôznym počiatkom spustenia reakcie, čo predstavuje zmiešanie dvoch roztokov. Tento problém bol vyriešený pomocou programu vytvoreného v MatLabe špeciálne pre túto prácu. Následne sa merala kinetika pri rôznych podmienkach, jednalo sa o zmenu koncentrácie RH421 a ATP. Pri meraní s rôznou koncentráciou RH421 boli dosiahnuté porovnateľné výsledky s literatúrou. Avšak meranie s rôznou koncentráciou ATP neposkytli výsledky odpovedajúce literatúre a toto meranie bude potrebovať ďalšie optimalizácie.The aim of this thesis was the isolation and optimization of kinetics measurements of Na+/K+-ATPase by stopped-flow method under different conditions. The theoretical part of the thesis is devoted to membrane transport, structure and mechanism of Na+/K+-ATPase. Subsequently, we focused on kinetics methods used for monitoring of enzyme kinetics. In the practical part, we described isolation and measurement of kinetics of Na+/K+-ATPase. The fluorescent dye RH421 was used for measurements. The change of fluorescence intensity was monitored after fast mixing two solutions, when one contained Na+/K+-ATPase incubated with the dye and the other only ATP solution. During the optimization of method few problems have occurred with the different starting time of the reaction, such as mixing solutions together. This problem was solved by program created in MatLab especially composed for this work. Afterward, we measured the kinetics under different conditions that means the alternation of concentration of RH421 and ATP. During the measurement with different concentration of RH421, comparable results with literature were achieved. However, measurement with different concentration of ATP failed to provide comparable results with literature and this measurement will need further optimization.