Low-frequency vibrations (0.5–5 Hz) that harm drivers occur in off-road vehicles. Thus, researchers have focused on finding methods to effectively isolate or control low-frequency vibrations. A novel nonlinear seat suspension structure for off-road vehicles is designed, whose static characteristics and seat-human system dynamic response are modeled and analyzed, and experiments are conducted to verify the theoretical solutions. Results show that the stiffness of this nonlinear seat suspension could achieve real zero stiffness through well-matched parameters, and precompression of the main spring could change the nonlinear seat suspension performance when a driver’s weight changes. The displacement transmissibility curve corresponds with the static characteristic curve of nonlinear suspension, where the middle part of the static characteristic curve is gentler and the resonance frequency of the displacement transmissibility curve and the isolation minimum frequency are lower. Damping should correspond with static characteristics, in which the corresponding suspension damping value should be smaller given a flatter static characteristic curve to prevent vibration isolation performance reduction.