Due to the lack of basic information on Chinese ghee and the increasing demand of consumers for natural oils, this study aims to explore and distinguish the quality characteristics of ghee in different regions of China. Ghee samples from 16 regions of Qinghai Province, Gansu Province, Xinjiang Uyghur Autonomous Region, and Tibet Autonomous Region were selected and their physicochemical properties, fatty acids, texture, antioxidant and antibacterial activities were determined. The results showed that: (1) The physicochemical properties of ghee were different from different regions, but the freshness and fat content are generally high. The results of iodine value and saponification value suggest that the fatty acid composition is good; (2) The unsaturated fatty acid/saturated fatty acid content of ghee in Tibet and Xinjiang ranges from 63.05% to 79.13%, which is better than that in other regions; (3) Gansu Diebu ghee has the highest hardness (40.69 N); (4) Ghee from different regions has good antioxidant activity, DPPH free radical scavenging activity is 30.45% to 58.06%, ABTS free radical scavenging activity is 41.14% to 65.53%, and has varying degrees of inhibition on gram-positive bacteria. In addition, yak ghee, cattle-yak ghee and cow ghee have better fatty acid composition and antibacterial ability than scalper ghee. The results of this study distinguish the differences in the quality characteristics of yak ghee in different geographical regions. Therefore, it can provide a theoretical basis for the origin tracing and quality-oriented improvement of yak ghee.