E- and P-selectin recognize a wide and overlapping range of oligosaccharide ligands including sialyl-Lewis X (sLeX) through their highly homologous C-type lectin domains. We report that an epitope apparently conserved between E- and P-selectin is functionally involved in ligand recognition although distantly located from the conventional carbohydrate binding site. We found that a previously established anti-E-selectin monoclonal antibody (mAb), 1.2B6, is cross-reactive with P-selectin, and that the 1.2B6 epitope is in the C-type lectin domain and identical to or overlapping with an epitope recognized by other independently established anti-E- and P-selectin dual-specific mAb. The epitope has been mapped by others to a region distant from the previously identified carbohydrate binding site of E-selectin in its three-dimensional structure. Nevertheless, it is of note that all dual-specific mAb, including 1.2B6, inhibited E- or P-selectin-mediated cell adhesion and also binding to sLeX. Engagement of the apparently conserved epitope by the dual-specific mAb may lead to inhibition of the ligand binding ability of E- and P-selectin by a previously uncharacterized mechanism(s) rather than by direct inhibition of sLeX binding to the hitherto identified ligand binding site.