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This study aimed to investigate the test-retest reliability of peak force in the isometric squat across the strength spectrum using coefficient of variation (CV) and intra-class correlation coefficient (ICC). On two separate days, 59 healthy men (mean (SD) age 23.0 (4.1) years; height 1.79 (0.7) m; body mass 84.0 (15.2) kg) performed three maximal effort isometric squats in two positions (at a 120° and a 90° knee angle). Acceptable reliability was observed at both the 120° (CV = 7.5 (6.7), ICC = 0.960 [0.933, 0.977]) and 90° positions (CV = 9.2 (8.8), ICC = 0.920 [0.865, 0.953]). There was no relationship between peak force in the isometric squat and the test-retest reliability at either the 120° (r = 0.052, p = 0.327) or 90° (r = 0.014, p = 0.613) positions. A subgroup of subjects (n = 17) also completed the isometric squat test at a 65° knee angle. Acceptable reliability was observed in this position (CV = 9.6 (9.3), ICC = 0.916 [0.766, 0.970]) and reliability was comparable to the 120° and 90° positions. Therefore, we deem isometric squat peak force output to be a valid and reliable measure across the strength spectrum and in different isometric squat positions.