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Agarose bound Phaseolus Vulgaris Erythroagglutinin (PHA-E)

Catalog Number: AL-1123

Availability: In stock

$145.00
Unit Size: 2 ml

Agarose bound Phaseolus Vulgaris Erythroagglutinin (PHA-E) Zoom

Overview

    Features:

    • Matrix is heat stable, cross-linked 4% agarose beads with a molecular exlusion of about 2x107 daltons
    • Bead diameter ranges in size from 45-165 microns
    • Matrix is stable in solutions at pH 3-11 as well as many organic solvents
    • Immobilized lectins are prepared using affinity purified lectins
    • Covalent attachment preserves lectin activity and minimizes conformational changes that might result in nonspecific or hydrophobic interactions
    • Hydrophilic spacer arm is inserted between the lectin and the matrix
    • Conjugated proteins are not leached off the beads by Tris or other routinely used buffers
    • No residual charges present after conjugation.  This minimizes non-specific binding to the matrix
    • Product supplied as a 1:1 suspension in buffer
    • 3 mg lectin/ml gel
    • Elution: 100 mM acetic acid or Glycoprotein Eluting Solution (ES-2100)

     

    Additional Information

    Catalog Number AL-1123-2
    Unit Size 2 ml
    Country of Manufacture United States
    Applications Glycobiology, Affinity Chromatography
    Matrix Conjugate Lectins
    Sugar Specificity Galactose, Complex Structures
    Conjugate Agarose
Additional Info

    Details

    Phaseolus vulgaris agglutinin is the name ascribed to a family of lectins, each of which consists of four subunits. There are two different types of subunits. One appears to be involved primarily in red cell agglutination and has been designated the E subunit (for erythroagglutinin). The other type is involved in lymphocyte agglutination and mitogenic activity and has been termed the L subunit (for leucoagglutinin). These subunits combine to produce five isolectins.

    One of these isolectins has four E subunits and is designated PHA-E. PHA-E possesses strong hemagglutinating activity but is a poor mitogen. PHA-L, with four L type subunits, does not agglutinate red cells but is a potent mitogen. The other three isolectins, designated E3L1, E2L2, and E1L3, have erythroagglutinating and mitogenic activities proportional to the number of respective E or L subunits. We have termed the mixture of the five isolectins PHA (E+L).

    PHA-L has been found to be an excellent, specific marker for use in anterograde neuronal tracing.

    Agarose bound* PHA-E 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.

    Elution: 100 mM acetic acid

    *3 mg lectin/ml gel

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