Context: This study aimed to examine the differences in electromyographic (EMG) activity of the biceps femoris long head (BFlh) and semitendinosus (ST) muscles, break-point angle (BPA), and the angle at peak BFlh EMG activity between bilateral and unilateral Nordic hamstring exercise (NHE) on a sloped platform. Design: This study was designed as a case-control study. Methods: Fourteen men participated in the study. The participants initially performed maximum voluntary isometric contraction (MVIC) on the prone leg curl to normalize the peak hamstring EMG amplitude as the %MVIC. Then, participants were randomized to perform the following 3 variations of NHE: bilateral (N40) or unilateral (N40U) NHE with a platform angle of 40°, and unilateral NHE with a platform angle of 50° (N50U). The EMG activities of the BFlh and ST and the knee flexion angle during the NHE variations were recorded to calculate the EMG activity of the BFlh and ST in terms of the %MVIC, the angle at peak BFlh EMG, and BPA. Results: The BFlh %MVIC was significantly higher in N40U (P < .05) and N50U (P < .05) than in N40. A significant difference in BFlh %MVIC and ST %MVIC was observed between N40U (P < .05) and N50U (P < .05). The mean values of BPA and the angle at peak BFlh EMG were <30° for all NHE variations. Conclusions: In the late swing phase of high-speed running, BFlh showed higher EMG activity; thus, unilateral NHE may be a specific hamstring exercise for hamstring injury prevention.