Succinylated Triticum vulgaris Lectin (Succ WGA) - Separopore® 4B

Succinylated Triticum vulgaris (Wheat) lectin (WGA) has been reported to have properties distinct from the native form. Evidence suggests that Succinylated Wheat Germ does not bind sialic acid, unlike the native form, but retains its specificity toward N-acetylglucosamine. Using conjugates of the native lectin and the succinylated form can provide a system to distinguish between sialylated glycoconjugates and those containing only N-acetylglucosamine structures.

Separopore 4B bound succinylated WGA is prepared using our affinity-purified lectins. Separopore 4B (4% Agarose) with a molecular weight exclusion limit of about 2x107 daltons are used as the solid-phase matrix to which the lectins are covalently coupled. The attachment of the lectins to the beads is carefully controlled to preserve lectin activity and minimize conformational changes of the bound lectins that might result in nonspecific ionic or hydrophobic interactions. The technique we have developed to couple lectins to agarose beads inserts a hydrophilic spacer arm between the lectin and the matrix.

This coupling method provides several advantages over the traditional cyanogen bromide procedure:

• Maximum carbohydrate binding activity of the coupled lectins is retained
• Linkage is stable over a range of pH values
• Conjugated proteins are not leached off the beads by Tris or other routinely used buffers
• No residual charges are present after conjugation. This minimizes non-specific binding to the matrix.

Our Separopore 4B bound lectins are supplied at a constant concentration of lectin per ml of settled beads. The concentration for each lectin is selected to achieve the highest glycoconjugate binding capacity per mg of lectin present in the beads.

Technical Specifications:

Ligand Source: Succinylated Triticum vulgaris

Matrix: Separopore 4B-CL (crosslinked agarose beads, 4%)

Particle size range: 52 - 165 µm

Matrix activation: Cyanogen bromide

Matrix attachment: Amino

Ligand density: 2 mg Wheat / mL of settled gel

Binding Capacity: 2-3 mg/mL

Inhibiting/Eluting Sugar: Chitin hydrolysate or GlcNAc with acid or salt

Carbohydrate for Elution: 0.2M Chitin hydrolysate or GlcNAc with acid or salt

Blood Group Specificity: A, B, O

Form: 50:50 Liquid Suspension, PBS (1x) pH 7.4 w/ 0.01% Thimerosal

pH stability: 4 - 9

Storage: 2 - 8 C, DO NOT FREEZE

Application note: Calcium is required for binding.