Implantation failure is one of the major causes of infertility and remains a major barrier to assisted reproduction success. Initial receptivity to implantation is regulated by the endometrial luminal epithelium under maternal hormonal control. Identification of epithelial cell surface components involved in embryo attachment will have translational applications in early pregnancy failure, infertility and contraception. In this study, vectorial biotinylation has been used to characterize the apical glycoproteome of Ishikawa cells, a polarized cell line that serves as a model of the implantation-receptive human endometrial luminal epithelium. Of 46 surface-associated glycoproteins detected by mass spectrometry, half are newly reported in this cell type; a subgroup of these were chosen for evaluation in tissue, and all were shown to be expressed apically in vivo in the mid-secretory (implantation) phase of the menstrual cycle, thus validating the model. Eleven adhesion molecules were detected, some already known to be involved in implantation, others novel. Cadherin 6, desmoglein 2 and plexin b2 were surprisingly found in the apical as well as the lateral membrane domain; their knock-down compromised epithelial integrity. This method of targeting glycosylated apical surface moieties in a polarized epithelial culture model shows excellent selectivity and identifies candidate cell adhesion molecules that are also present in vivo in secretory phase endometrial epithelium.