The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use 2.5µl for 106 cells. ab106163-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Use a concentration of 10 - 20 µg/ml.
Sialyl-Tn is a carbohydrate antigen overexpressed in several epithelial cancers including breast cancer, and usually associated with poor prognosis. Sialyl-Tn is synthesized by a CMP-Neu5Ac: GalNAc alpha2,6-sialyltransferase: ST6GalNAc I, which catalyzes the transfer of a sialic acid residue in alpha2,6-linkage to the GalNAcalpha1-O-Ser/Thr structure. The resulting disaccharide (Neu5Acalpha2-6GalNAcalpha1-O-Ser/Thr) cannot be further elongated and sialyl-Tn expression results therefore in a shortening of the O-glycan chains
Overlay histogram showing DU145 cells stained with ab76756 (red line). The cells were fixed with 80%methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76756, 0.5µg/1x106 cells) for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 (1µg/1x106 cells ). Acquisition of >5,000 events was performed.