Wisteria floribunda (WFA, Japanese Wisteria) (Agarose)

Wisteria Floribunda lectin is a 116,000 to 125,000 molecular weight glycoprotein with an isoelectric point of pH 5.4. Two subunits of about 60,000D are produced in SDS gels, dissociating into 4 chains of about 30,000D in the presence of reducing agents. The binding specificity is not completely clear but this lectin appears to preferentially bind carbohydrate structures terminating in N-acetylgalactosamine linked alpha or beta to the 3 or 6 position of galactose. Used to fractionate lymphocyte populations, the lectin although not mitogenic, elicits the production of lymphokines from murine splenocytes. Agarose bound Wisteria Floribunda Lectin 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 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: >4mg N-acetylgalactosaminyl glycoprotein/ml gel, Inhibiting/Eluting Sugar: 200mM N-acetylgalactosamine 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.