‘Sit-to-stand’ exercise uses the repetitive motion of standing up and sitting down in a chair, a common activity of daily living. A new assessment using an incremental sit-to-stand exercise test employs an external sound to control the speed of standing-up and allows increases in work rate. The aims of the study were to examine the effect of different seat heights on peak oxygen uptake (peak VO₂) during an incremental sit-to-stand exercise and to assess any difference between peak VO₂ values during incremental sit-to-stand exercise compared with a cycle ergometer test. Thirteen healthy young women (age: 23.1 ± 2.6 years, height: 1.61 ± 0.06 m, body mass: 51.9 ± 7.4 kg·m^-2) participated in four incremental sit-to-stand tests with different seat heights and cycle tests in random order. The seat heights were adjusted to 100%, 80%, 120%, and 140% of knee height distance (100%, 80%, 120%, and 140% incremental sit-to-stand exercise, respectively). The peak VO₂ and completion time were measured during incremental sit-to-stand and cycle ergometer tests, and repeated-measures analysis of variance and Student’s paired t-test with Holm’s method were used to evaluate differences between these variables. The peak VO₂ values increased by about 10-12 mL·min^-1·kg^-1 as the seat height on the ISTS decreased over a 60% range of lower leg lengths. The peak VO₂ values on the 80%, 100%, 120%, and 140% incremental sit-to-stand tests were about 11%, 25%, 40%, and 50% lower than that on the cycle ergometer test, respectively. The peak VO₂ on the incremental sit-to-stand test increased as seat height decreased. These findings are useful to determine which seat height on the incremental sit-to-stand tests test is suitable for different populations.