The purpose of the study is to examine the effect of the ankle joint range of motion (ROM) on the vertical jump (VJ) performance of adult handball players. The active (ACT) and passive (PAS) ankle joint ROM of 12 male members of the U21 National Handball Team with the knee joint at 0°, 40°, and 90° flexion (0° = fully extended knee) was evaluated using a video analysis measuring method. Participants also performed maximum VJ with (CMJ) and without (SQJ) countermovement, as well as with (AS) and without (NAS) an arm swing. Statistical analyses included 2 × 2 × 3 MANOVA, 2 × 2 repeated measures ANOVA, and Pearson’s correlation. Results reveal that PAS-ROM was larger (p < 0.05) in all knee joint flexion angles. ROM was smaller (p < 0.05) by approximately 10° at 0° compared to 90° knee flexion. No lateral effects on ROM due to the handedness of the players were observed. AS and CM resulted in increased jump height (p < 0.05). Finally, ACT-ROM when the knee joint was flexed at 40° was highly correlated (r ≥ 0.66, p < 0.05) with VJ performance except for CMJ-AS. In conclusion, the differences in the bi-articular gastrocnemius muscle flexibility due to the alteration of the angular position of the examined joints affected the ability to generate impulse during the VJ tests.