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PNGase F treatment on immunoprecipitated CEA demonstrated the presence of SLex in N-glycans. CEA from both cell lines is heavily N-glycosylated as PNGase F mediated deglycosylation shifts the SDS-PAGE migration of CEA from 150-200 kDa towards its predicted molecular weight of around 71 kDa. The PNGase F treated CEA from MST3Gal IV cells was negative in the SLex western blot analysis confirming that the SLex epitope is present on CEA N-glycans (black arrows).
