Concanavalin A (Jackbean) Lectin (Con A) - Separopore® 4B-CL Columns

Applications:

Con A-Separopore 4B-CL is used for the purification and enrichment of glycoproteins, polysaccharides and glycolipids as it binds molecules containing α-D-mannopyranosyl, α-D-glucopyranosyl and sterically related residues. Con A-Separopore 4B-CL has also be used in other application areas including purification of enzyme-antibody conjugates, purification of IgM and separation of membrane vesicles. Con A-Separopore 4B-CL is ideal for isolation of cell surface glycoproteins from detergent solubilized membranes.Concanavalin A is one of the most widely used and well characterized lectins. Con A has broad applicability primarily because it recognizes a commonly occurring sugar structure, a-linked mannose. Since a wide variety of serum and membrane glycoproteins have a “core oligosaccharide” structure which includes a-linked mannose residues, many glycoproteins can be examined or purified with Con A and its conjugates. Briefly, Con A has been utilized in hormone receptor studies, mitogenic assays, characterization of normal and malignant cells, glycoprotein purification, viral antigen isolation, dextran and mannan fractionation, cell agglutination studies, bacterial aggregation, membrane fluidity and lateral mobility investigations, turbidimetric assays for sugars, lymphokine production, as well as in many other applications.

Separopore conjugated Concanavalin A is prepared from affinity-purified lectin. Heat stable, cross-linked 4% agarose beads with a molecular weight exclusion limit of about 2 x 10⁷ 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. Mn⁺⁺ and Ca⁺⁺ (1 mM each) should be present in the binding buffers.

Technical specifications

• Ligand: Concanavalin A (Con A)
• Matrix: Separopore 4B-CL (crosslinked agarose, 4%)
• Particle size range: 52 - 165 µm
• Matrix activation: Cyanogen bromide
• Ligand density: > 10 mg Concavalin A / ml drained gel
• Binding capacity: > 25 mg HRP / ml drained gel
• Exclusion limit: approximately 2 x 10⁷
• Inhibiting/Eluting Sugar: 200 mM α-methyl mannoside/200 mM α-methyl glucoside mixture
• Storage: 2 - 8 C
• Supplied as suspension in 0.1M sodium acetate pH 6, 1M NaCl, 1mM CaCl₂, 1mM MnCl₂ with 20% ethanol

References:

• Purification and partial characterisation of pectin methylesterase produced by Fusarium asiaticum. Fungal Biol. (2011) 115: 1112-21.
• Echinococcus multilocularis: purification and characterization of glycoprotein antigens with serodiagnostic potential for canine infection. Exp Parasitol. (2011) 128: 50-6.
• Improved secretion of human Fas ligand extracellular domain by N-terminal part truncation in Pichia pastoris and preparation of the N-linked carbohydrate chain trimmed derivative. Protein Expr Purif. (2008) 60: 205-13.
• Analysis of lectin-bound glycoproteins in snake venom from the Elapidae and Viperidae families. J Proteome Res. (2004) 3: 383-92.
• Cytokinin oxidase activity and cytokinin content in roots of sunflower under water stress. Indian J Exp Biol. (2001) 39: 786-92.
• Glycome project: concept, strategy and preliminary application to Caenorhabditis elegans. Proteomics. (2001) 1: 295-303.
• Purification of alpha-amylase isoenzymes from Scytalidium thermophilum on a fluidized bed of alginate beads followed by concanavalin A-agarose column chromatography. Protein Expr Purif. (2000) 20: 162-8.
• Bovine spleen cathepsin A: characterization and comparison with the protective protein. J Biochem. (1998)123: 701-6.
• The production and purification of functional decorin in a baculovirus system. Biochem Biophys Res Commun. (1997) 232: 91-5.
• Roles of heme iron-coordinating histidine residues of human hemopexin expressed in baculovirus-infected insect cells. Proc Natl Acad Sci U S A. (1994) 91: 8423-7.
• SS33410, an inhibitor for inflammation, blocks the intracellular transport of VSV G glycoprotein in BHK cells.Biochem Biophys Res Commun. (1994) 199: 1073-80.
• Identification of a 130-kilodalton human biliary concanavalin A binding protein as aminopeptidase N.Gastroenterology. (1994) 106: 755-62.
• Identification by microsequencing of lipopolysaccharide-induced proteins secreted by mouse macrophages. J Immunol. (1993) 151: 1535-47.
• cDNA cloning of guinea pig monocyte chemoattractant protein-1 and expression of the recombinant protein. J Immunol. (1993) 150: 5025-32.
• Rapid purification and N-terminal sequencing of a potato tuber cyclic nucleotide binding phosphatase. Biochim Biophys Acta. (1992) 1159: 179-84.
• A partial characterization of a Sertoli cell-secreted protein stimulating Leydig cell testosterone production.Endocrinol Jpn. (1992) 39: 209-15.
• Receptor glycosylation regulates the affinity of histamine H1 receptors during smooth muscle cell differentiation. Mol Pharmacol. (1989) 35: 311-8.
• Active suppression of host-vs graft reaction in pregnant mice. IX. Soluble suppressor activity obtained from allopregnant mouse decidua that blocks the cytolytic effector response to IL-2 is related to transforming growth factor-beta. J Immunol. (1988) 141: 3833-40.
• Characterization of fucosyl oligosaccharides associated with synaptic membrane and synaptic junctional glycoproteins. J Neurochem. (1987) 48: 1604-11.
• Lectin affinity and PAGE analysis of soluble axonally transported glycoconjugates in the rat visual system.Brain Res. (1986) 399: 42-50.

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