UEA-I-Biotin, Conjugated

Histochemistry 1: 100-1:1000; WB 1:200-1:2,000; ELISA 1:500-1:2,500. Dilute biotinylated lectin in PBS containing 0.1 mM calcium chloride ions. For Histochemistry the tissues are processed same as for Immunohistochemistry, after blocking step biotinylated lectin is applied followed by streptavidin conjugated to enzyme and chromogen. For WB incubate membrane with blocking protein followed by biotinylated lectin, streptavidin enzyme conjugate and chromogen. For ELISA the target proteins are absorbed on ELISA plate at a concentration of 5-10 µg/ml (50-100 µl), followed by blocking with protein solution, Biotinylated lectin, Streptavidin enzyme conjugate, ELISA substrate. Please refer to Histochemistry, WB and ELISA protocol for detail information.

The anti-H (O) hemagglutinating activity of Ulex europaeus has been used widely to confirm blood group O activity. UEA I binds to many glycoproteins and glycolipids containing α –linked fucose residue. The purified lectin appears to be a dimer of two distinct polypeptide chains associated by noncovalent forces. This lectin does not react with Lea active blood group substance. The native protein has pI=6.0-6.1 and exhibits a Molecular weight 60,000-68,000 during gel filtration on Sephadex column.