Soybean Agglutinin (SBA, Glycine max) (Agarose)

Composed of four subunits of approximately equal size, soybean agglutinin is a family of closely related isolectins. This glycoprotein has a molecular weight of about 120,000 and an isoelectric point near pH 6.0. SBA preferentially binds to oligosaccharide structures with terminal a- or b-linked N-acetylgalactosamine, and to a lesser extent, galactose residues. Binding can be blocked by substitutions on penultimate sugars, such as fucose attached to the penultimate galactose in blood group B substance. SBA has been used in glycoprotein fractionation, histochemical applications and cell sorter analysis. An important application for soybean agglutinin is the separation of pluripotential stem cells from human bone marrow. Cells fractionated by SBA do not produce graft vs host disease and can be used in bone marrow transplantation across histocompatibility barriers. It should be noted that some forms of SBA seem to be excellent in separating human cells while others are better for cells of other species. Agarose bound Soybean Agglutinin is prepared from affinity-purified lectin. Heat stable, cross-linked 4% agarose beads with a molecular weight exclusion limit of about 2x10e7 are used as the solid-phase matrix to which the lectin is covalently bound. The attachment of the lectin to the solid phase is carefully controlled in order to preserve the activity of the lectin as well as to minimize conformational changes of the bound lectin which might result in nonspecific ionic or hydrophobic interactions. The technique we have developed to couple lectins to agarose provides a very hydrophilic spacer arm between the protein and the matrix. This ensures maximum expression of the carbohydrate binding activity of the lectin. The linkage is very stable over a range of pH values and, unlike cyanogen bromide linkages, proteins are not leached off the gel by Tris or other routinely used buffers. In addition, residual charges generated during cyanogen bromide conjugation which can produce nonspecific binding are not present on the gel following our coupling procedure. \n\nApplications: \nSuitable for use in ELISA. Other applications not tested.\n\nRecommended Dilution:\nOptimal dilutions to be determined by the researcher.\n\nBinding Capacity:\n4.8mg GalNAc-BSA/ml of gel\n\nInhibiting/Eluting Sugar: \n200mM N-acetylgalactosamine 4mg lectin/ml gel\n\nActivity:\n2x10e7\n\nStorage and Stability:\nMay be stored at 4 degrees C. For long-term storage, aliquot and store at 4 degrees C. Do not freeze. Aliquots are stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.

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