Podoplanin (PDPN)/Aggrus is a type I transmembrane O-glycoprotein, which is expressed in several normal tissues including podocytes of kidney and lymphatic endothelial cells. PDPN activates platelet aggregation by binding to C-type lectin-like receptor-2 (CLEC-2) on platelet; however, only bovine PDPN (bovPDPN) does not possess the platelet-aggregating activity. Although many monoclonal antibodies (mAbs) against human PDPN, mouse PDPN, rat PDPN, and rabbit PDPN have been established, anti-bovPDPN mAbs have not been developed. In this study, we immunized mice with the recombinant proteins of bovPDPN and developed anti-bovPDPN mAbs, which are useful in immunohistochemical analysis. One of the clones, PMab-44, is useful for detecting podocytes and lymphatic endothelial cells in normal bovine tissues. PMab-44 also detected bovPDPN specifically in flow cytometry. PMab-44 is expected to be useful for investigating the function of bovPDPN.