Objectives: The aim of this study was to investigate the effect of immersion in sodium hypochlorite (NaOCl) on cyclic fatigue resistance of three different rotary nickel–titanium (NiTi) retreatment files. Material and methods: A total of 90 new ProTaper, D-RaCe, and Mtwo retreatment files were tested. Thirty files of the same brand were randomly assigned to two groups (n = 15). Group 1 was no immersion (control group) and 16 mm of group 2 instruments were immersed in 5 % NaOCl at 37 °C for 5 min. All instruments were then tested for cyclic fatigue. Resistance to cyclic fatigue was determined by counting the numbers of cycles to failure in a 60° curve with a 5-mm radius, recording the time in seconds to fracture. Data were analyzed by two-way analysis of variance. Results: Resistance to cyclic fatigue decreased significantly for D-RaCe retreatment files after immersion in NaOCl. ProTaper and Mtwo retreatment files were not affected from immersion to NaOCl. D-RaCe retreatment instruments showed better cyclic fatigue resistance than ProTaper and Mtwo retreatment instruments, and the difference was statistically significant (p < 0.05). There was no significant difference between ProTaper and Mtwo retreatment instrument groups (p > 0.05). Conclusions: D-RaCe retreatment instruments had the highest cyclic fatigue resistance among retreatment files tested in this study, but immersion to NaOCl decreased the cyclic fatigue resistance of D-RaCe retreatment instrument. Clinical relevance: It should be considered that cyclic fatigue resistance of D-RaCe can decrease in contact with NaOCl during the removal of canal filling material.