100Cr6 steel is one of the most widely used bearing steels and a representative of first-generation bearing steel. Many engineering applications require rolling bearings to run at a high temperature. Therefore, it is necessary to improve the high temperature properties of 100Cr6 steel. In this paper, the effect of Nb on high temperature dry tribological behavior, including worn surface and friction coefficient, was analyzed by a wear test when Nb content was 0.018% and 0.040%. The results show that the microstructure is refined gradually, the hardness is improved, and wear volume decreases by 31.8% at most with the increase of Nb content. At 50 °C, the friction coefficient of 100Cr6 steel can be reduced by adding a small amount of Nb, but this effect will be weakened if the content of Nb is too high. In addition, excess Nb increases the hard precipitation of NbC, which aggravates the abrasive wear and leads to the increase in the depth of the worn surface. At 125 °C, the effect of Nb on tribological properties is weaker. With the increase of temperature, the steel substrate softens, and the oxide particles increase, which aggravates the abrasive wear and oxidation wear and makes the wear volume increase significantly.