Galanthus nivalis (Snowdrop Bulb) (GNA) (Agarose)

Galanthus nivalis lectin is a small molecular weight tetramer consisting of subunits of about 13,000D. It has an isoelectric point below pH 4.6. The lectin contains little or no carbohydrate and, unlike most mannose-specific lectins, is not a metalloprotein and does not require Ca++ or Mn++ for binding. Binding seems to be preferentially directed toward structures containing (a-1,3) mannose residues. Also in contrast to most mannose-binding lectins, GNL will not bind alpha linked glucose. Reports indicate that this lectin binds IgM but not IgG of the immunoglobulin classes of rat and mouse. The only protein from human serum reported to bind to this lectin is a2-macroglobulin. Agarose bound Galanthus Nivalis 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 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. Binding Capacity: 3mg mannose-containing serum protein/ml of gel, Inhibiting/Eluting Sugar: 100mM-200mM a-methyl mannoside 3mg lectin/ml gel, Activity: 2x10e7, Applications: Suitable for use in ELISA. Other applications not tested. Recommended Dilution: Optimal dilutions to be determined by the researcher. 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.