Background: Total arterial occlusive pressure (AOP) is used to prescribe pressures for surgery, blood flow restriction exercise (BFRE) and ischemic preconditioning (IPC). AOP is often measured in a supine position; however, the influence of body position on AOP measurement is unknown and may influence level of occlusion in different positions during BFR and IPC. The aim of this study was therefore to investigate the influence of body position on AOP. Methods: Fifty healthy individuals (age = 29 ± 6 y) underwent AOP measurements on the dominant lower-limb in supine, seated and standing positions in a randomised order. AOP was measured automatically using the DelfiPersonalised Tourniquet System device, with each measurement separated by 5 min of rest. Results: Arterial occlusive pressure was significantly lower in the supine position compared to the seated position (187.00 ± 32.5 vs 204.00 ± 28.5 mmHg, p < 0.001) and standing position (187.00 ± 32.5 vs 241.50 ± 49.3 mmHg, p < 0.001). AOP was significantly higher in the standing position compared to the seated position (241.50 ± 49.3 vs 204.00 ± 28.5 mmHg, p < 0.001). Discussion: Arterial occlusive pressure measurement is body position dependent, thus for accurate prescription of occlusion pressure during surgery, BFR and IPC, AOP should be measured in the position intended for subsequent application of occlusion.