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Featuring:
Plus
quality reagents for:
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| Agarose bound* Concanavalin A (Con A)
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Con A recognizes α-linked mannose present as part of a “core
oligosaccharide” in many serum and membrane glycoproteins.
At neutral and alkaline pH, Con A exists as a tetramer of
four identical subunits; below pH 5.6, Con A dissociates into
active dimers of 52 kDa. Acetylation, succinylation, or other
derivatizations can also produce stable forms with dimeric
structures. (See succinylated Con A). “Nicks” in the sequence
are often present in the purest preparations due to hydrolytic
damage within the seeds.
Con A requires calcium or manganese ions at each of its four
saccharide binding sites. Although these divalent metal ions
are bound tightly to the polypeptide structure, buffers which
can bind calcium (such as phosphate) generally should be
avoided in diluting Con A, since a gradual loss in activity may
occur.
Agarose bound* Con A is 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.
Inhibiting/Eluting Sugar: mixture of 200 mM α-methylmannoside/200 mM
α-methylglucoside
*6 mg lectin/ml gel
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