Syngeneic breast tumors were developed by inoculating of 4T1 cells, into the left flank of female Balb/c mice. When tumor volume reached 200-250 mm3, mice were selected randomized way to receive the treatment of anti-TIM-3 monoclonal antibody (mAb) (Clone: RMT3-23, BioXcell, cat. # BE0115, RRID: AB_10949464). Anti-TIM-3 mAb (250 µg/injection) was administered via the intraperitoneal (i.p.) route. Primary tumors were allowed to developed for up-to 40 days. Primary tumor histology, liver metastasis, and serum cytokine and tumor tissue proteome were analysed and compared between the control tumor and TIM-3 antibody treated tumor.