This study used a mechanical test to evaluate the flexibility of instruments from the K3 (conicity 0.04) and the ProTaper Universal systems when they were new and after 5 uses in simulated canals. Five sets of instruments of each system were tested: K3 (15, 20, 25, 30, 35, 40 and 45) and ProTaper Universal (S1, S2, F1, F2, F3, F4 and F5). Each set of instruments was used to prepare a simulated canal and the same set of instruments was used 5 times (50 canals). The number of each subgroup represented the number of uses: 0 (control), 1, 3 and 5 uses. Before and after each use, the instruments were submitted to a mechanical flexibility test performed in a Versat 502 universal testing machine. Interactions between the instrument and the number of uses were analyzed by ANOVA and Tukey's test at a 5% level of significance. Instruments from both systems presented lower flexibility after the third use compared to the flexibility obtained after uses 0 and 1 (p<0.05), and maintained the same flexibility after the fifth use. The flexibility of instruments from the K3 system decreased with the increase of diameter, irrespective of the number of uses. Among the instruments from the ProTaper Universal system, the shaping files presented greater flexibility than the finishing files. F2 and F3 were the least flexible instruments, and F4 and F5 presented flexibility values similar to those of F1.