CONTEXT Individuals with anterior cruciate ligament reconstruction (ACLR) have quadriceps dysfunction that contributes to physical disability and posttraumatic knee osteoarthritis. Quadriceps function in the ACLR limb is commonly evaluated relative to the contralateral uninjured limb. Bilateral quadriceps dysfunction is common in individuals with ACLR, potentially biasing these evaluations. OBJECTIVE To compare quadriceps function between individuals with ACLR and uninjured control participants. DESIGN Cross-sectional study. SETTING Research laboratory. PATIENTS OR OTHER PARTICIPANTS Twenty individuals with unilateral ACLR (age = 21.1 ± 1.7 years, mass = 68.3 ± 14.9 kg, time since ACLR = 50.7 ± 21.3 months; females = 14; Tegner Score = 7.1 ± 0.3; 16 patellar tendon autografts, 3 hamstrings autografts, 1 allograft) matched to 20 control participants (age = 21.2 ± 1.2 years, mass = 67.9 ± 11.3 kg; females = 14; Tegner Score = 7.1 ± 0.4) on age, sex, body mass index, and Tegner Activity Scale. MAIN OUTCOME MEASURE(S) Maximal voluntary isometric knee extension was performed on an isokinetic dynamometer. Peak torque (PT), rate of torque development (RTD), electromyographic (EMG) amplitude, central activation ratio (CAR), and hamstrings EMG amplitude were assessed during maximal voluntary isometric knee extension and compared between groups using independent-samples t tests. Relationships between hamstrings co-activation and quadriceps function were assessed using Pearson correlations. RESULTS Participants with anterior cruciate ligament reconstruction displayed lesser quadriceps PT (1.86 ± 0.74 versus 2.56 ± 0.37 Nm/kg, P = .001), RTD (39.4 ± 18.7 versus 52.9 ± 16.4 Nm/s/kg, P = .03), EMG amplitude (0.25 ± 0.12 versus 0.37 ± 0.26 mV, P = .04), and CAR (83.3% ± 11.1% versus 93.7% ± 3.2%, P = .002) and greater hamstrings co-activation (27.2% ± 12.8% versus 14.3% ± 3.7%, P < .001) compared with control participants. Correlations were found between hamstrings co-activation and PT (r = -0.39, P = .007), RTD (r = -0.30, P = .03), and EMG amplitude (r = -0.30, P = .03). CONCLUSIONS Individuals with ACLR possessed deficits in PT, RTD, and CAR compared with control participants. Peak torque is the net result of all agonist and antagonist activity, and lesser PT in individuals with ACLR is partially attributable to greater hamstrings co-activation.