This lectin is a tetrameric glycoprotein composed of four subunits of nearly identical size. Each 30,000D subunit has a binding site for alpha- or beta-linked N-acetylglucosamine residues. Unlike other N-acetylglucosamine specific lectins, increasing the number of N-acetylglucosamine residues beyond two does not improve affinity. GSL II has been reported to be unique in its ability to recognize exclusively alpha- or beta-linked N-acetylglucosamine residues on the nonreducing terminal of oligosaccharides. Agarose bound Griffonia (Bandeiraea) Simplicifolia Lectin II is prepared from affinity-purified lectin. Heat stable, cross-linked 4% agarose beads with a molecular weight exclusion limit of about 2x107 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 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.
Applications:Suitable for use in ELISA. Other applications not tested.
Recommended Dilution:Optimal dilutions to be determined by the researcher.
Binding Capacity:≥4.0mg of N-acetylglucosaminyl glycoprotein/ml gel
Inhibiting/Eluting Sugar:Chitin Hydrolysate or 200mM N-acetylglucosamine 3mg lectin/ml gel
Activity:2x10e7
Storage and Stability:May be stored at 4°C. For long-term storage, aliquot and store at 4°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.
