Wisteria floribunda (WFA, Japanese Wisteria) (Biotin)

This 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. This biotinylated lectin conjugate is prepared from affinity-purified lectin and is optimally labeled with biotin. Essentially free of inactive lectin conjugate and containing no free biotin, this biotinylated lectin provides an ideal intermediate for examining glycoconjugates using the Biotin-Avidin System. First the biotin-labeled lectin is added, followed by ABC Reagent, Avidin D conjugate, or streptavidin derivative. Another possible application is in the isolation of lymphokines and other products of mitogenic stimulation. Applications: Suitable for use in ELISA, Western Blot and Immunohistochemistry. Other applications not tested. Recommended Dilution: Optimal dilutions to be determined by the researcher. Inhibiting/Eluting Sugar: 200mM N-acetylgalactosamine, Storage and Stability: Lyophilized powder may be stored at -20°C. Stable for 12 months at -20°C. Reconstitute with sterile ddH2O. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.