The 30 second Wingate Anaerobic Test (WAnT) is the gold standard measure of anaerobic performance. The present investigation aimed to determine if a previous WAnT using one body region significantly affected a subsequent WAnT using a different body region. Twelve male university students (n = 12, 23 ± 2 years, 84 ± 16.1 kg, 178.5 ± 7.4 cm) volunteered to complete two repeated WAnT protocols (either lower body WAnT followed by an upper body WAnT or vice versa) on two separate testing occasions. The upper body WAnT was conducted on a modified electromagnetically braked cycle ergometer using a flywheel braking force corresponding to 5% bodyweight. The lower body WAnT was conducted on an electronically braked cycle ergometer using a flywheel braking force corresponding to 7.5% bodyweight. Participants had a 1 minute rest period for transition between WAnTs. Data are reported as mean ± standard deviation. No significant differences were identified in power indices for the lower body between 30 s WAnTs. When the upper body WAnT was performed 2nd, absolute peak power (p < 0.01), mean power (p < 0.001) and relative mean power (p < 0.001) were significantly lower compared to when the upper body WAnT was performed 1st. The value of maximum revolutions per minute was significantly lower (p < 0.001) when the upper body WAnT was performed after the lower body WAnT, compared to when it was performed 1st (193.3 ± 11.4 1st vs 179.8 ± 14.4 2nd). Previous upper body sprint exercise does not significantly affect lower body sprint exercise; however, previous lower body sprint exercise severely compromises subsequent upper body sprint performance.