The aim of this study was to investigate the effect of quantification method on estimates of voluntary quadriceps muscle activation. Twenty‐two people with no history of serious lower extremity injuries underwent voluntary quadriceps activation testing at 60° of knee flexion. Estimates of quadriceps activation were derived with: (1) a formula based on the interpolated twitch technique (ITT); (2) the central activation ratio (CAR); and (3) a modified central activation ratio. Predictive equations were developed that describe the relationships between the three methods. Significant differences (P < 0.001) were observed between the estimates of voluntary quadriceps muscle activation obtained using the three methods (ITT percent activation = 93.0 ± 6.4%, CAR = 95.9 ± 3.8%, modified CAR = 98.5 ± 4.1%). Excellent correlation (r = 0.995) was observed between ITT‐based percent activation and the CAR method. The associations between these methods and the modified CAR approach were weaker. Quantification method affects activation estimates. The equations developed will assist scientists in accurately comparing the results of studies that use different methods of quantifying activation. Muscle Nerve, 2010