BACKGROUND: Blood group antigens are physiologically important differentiation markers in embryogenesis and development. Monoclonal antibody (MoAb) B2 recognizes a transient antigen expressed on late sympathoadrenal neuroendocrine precursors and early sympathetic neuroblasts. It has been suggested that MoAb B2 may recognize a globo-series glycosphingolipid (GSL) related to the P blood group family. STUDY DESIGN AND METHODS: MoAb B2 and two anti-LKE MoAbs, MC813-70 and RM1, were screened against a panel of GSL standards and isolated red blood cell (RBC) GSLs by high-performance thin layer chromatography (HPTLC) immunostaining. The ability of all three MoAbs to bind intact RBCs and two LKE+ renal cell carcinoma cell lines (A498, ACHN) were examined by flow cytometry and hemagglutination. RESULTS: MoAbs B2, MC813-70, and RM1 all specifically recognized monosialogalactosylgloboside (MSGG) on HPTLC immunostaining. Only MoAb MC813-70 bound intact RBC by flow cytometry and hemagglutination. Differential staining was observed between the three antibodies and two renal cell carcinoma cell lines. CONCLUSION: MoAb B2 recognizes MSGG or LKE antigen, suggesting that LKE may play a role in neuroendocrine differentiation from neural crest cells. Although MoAb B2 is not suitable for RBC phenotyping, it may be a useful immunologic reagent for the identification of human embryonic stem cells and renal cell and embryonic carcinoma.