Magnetorheological (MR) fluids are known for their controllable characteristics under the influence of magnetic fields and, hence, widely used as semi-active actuators for vibration control. Regardless of advantages such as fast response time and reversible property, MR fluids inevitably experience sedimentation caused by significant density mismatches between magnetic particles and carrier liquids. Moreover, the effect of the temperature on actuating characteristics is also one of the problems to be resolved for practical implementation. This study experimentally investigates the sedimentation behavior under various temperatures ranging from 25 to 70 °C using a multiguide-arm magnetic device that generates a uniform magnetic flux density across MR fluids. The sedimentation stability is then observed after 168 h at current inputs of 0, 1, and 2 A, respectively. Subsequently, the field-dependent rheological properties of MR fluids are evaluated using a rheometer and discussed, showing actuating capability, which depends on the viscosity, shear stress, and yield stress before (initial state) and after the sedimentation (sedimentation state). The field-dependent yield stresses, which directly represent the actuating force of the semi-active actuator, are specifically evaluated. Under the on-state condition (2 A) at a temperature of 70 °C, the yield stress decreased from 2.747 kPa (initial state) to 2.352 kPa (sedimentation state). By using this yield stress, the field-dependent damping force was evaluated, showing a decrement from 1672 N (initial state) to 1623 N (sedimentation state) at a velocity of 0.8 m/s. It is shown that the temperature causes the reduction of the actuating properties after the long-term operation. The insightful findings achieved in this work will provide useful information for the evaluation of actuating characteristics of smart MR fluids and the design of MR application systems subjected to particle sedimentation and temperature variation.