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Featuring:
Plus
quality reagents for:
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Agarose bound Lectins |
Lectin affinity chromatography is a simple and widely used
technique for the isolation of a variety of glycoconjugates. The
glycoconjugate is allowed to bind to the immobilized lectin,
and the unbound residual material is removed by washing.
The bound glycoconjugates are generally eluted with a solution
of a sugar known to inhibit binding of the particular lectin.
Soluble glycoproteins, hormones, antigens, polysaccharides,
detergent-solubilized membrane-bound glycoconjugates, cell
surface receptors, blood group substances, viral glycoproteins,
histocompatibility antigens, lymphokines, enzymes, lymphocyte
markers, serum proteins, and oncofetal antigens are only a few
of the substances that have been purified using immobilized
lectins.
Our immobilized lectins are prepared using our affinity-purified
lectins. Heat stable, cross-linked 4% agarose beads 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 agarose 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. Each lot is tested for its binding capacity using
glycoproteins known to bind the lectin. This provides a
guideline for the user and assures the quality of our agarose
bound lectins.
Glycoprotein Eluting Solutions
Vector Laboratories has developed five Glycoprotein Elution Solutions in the neutral pH range that maximize the yield of eluted glycoproteins and preserve the activity of the agarose-bound lectins for repeated use. All components of these ready-to-use Glycoprotein Eluting Solutions can be subsequently removed by dialysis.
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