Coatings are normally employed to meet some functional requirements. There is a kind of self-generated coating during use, such as the transfer layer during sliding, which may greatly affect the tribological behavior. Although the transfer layer has aroused much attention recently, the formation of the transfer layer closely depends on the service conditions, which need to be further studied. In this paper, the effects of sliding speed, normal load, and duration of wear test on the transfer layer thickness during friction of Ni/PTFE (Polytetrafluoroethylene) composites were experimentally investigated. The formation mechanism of transfer layer and the relationships between tribological properties and transfer layer thickness were analyzed in detail. It was found that the transfer layer thickness increased with increases of sliding speed and normal load; and after a period of wear test, the transfer layer thickness remained stable. The transfer layer thickness correlates linearly with the friction coefficient and wear volume of the PTFE composites. With the increase of the transfer layer thickness, the friction coefficient decreased, while the wear volume increased, which means that a uniform, thin, and stable transfer layer is beneficial for the reduction of friction and wear of the polymeric composites.