The AminoLink Plus Immobilization Kit contains AminoLink Plus Coupling Resin and additional reagents needed to perform 5 protein immobilizations, with expected yields of 2 to 40 mg protein per column. The AminoLink Plus Coupling Resin uses aldehyde-activated agarose beads for high-yield covalent coupling of antibodies (proteins) via primary amines to prepare columns for affinity purification.
Features of AminoLink Plus Coupling Resin:
• AminoLink Plus CouplingResin—aldehyde-activated crosslinked 4% beaded agarose • Ideal for antibodies and other proteins— immobilize molecules via primary amines (-NH2) • Flexible coupling conditions—efficient (>85%) coupling over a wide range of pH (4-10) and buffer conditions (PBS or other non-amine buffer with or without organic solvent); regular (PBS, pH 7.2) and enhanced (citrate-carbonate, pH 10) coupling protocols provided • Stable, permanent immobilization—Coupling reaction esults in stable, leak-resistant secondary amine bond between resin and ligand • Better than immobilization to CNBr-activated agarose—bond is more stable and uncharged, resulting in less nonspecific binding in affinity purification procedures • Versatile and reusable—prepared affinity resin is adaptable to column and batch affinity techniques and the resin is reusable for typical applications based on protein binding interactions
AminoLink Plus Resin is beaded agarose support that has been modified to contain aldehyde groups. Antibodies, other proteins, or any molecules with primary amines can be conjugated to the resin by an optimized form of reductive amination. Once an antibody or other ligand is immobilized, the prepared affinity resin can be used for a variety of affinity purification methods involving batch or column chromatography. The resin and linkage are stable in binding and elution conditions typically used in affinity chromatography, enabling prepared resin to be used for at least 10 rounds of affinity purification.
Specifications: Product Details: The AminoLink Plus Coupling Reaction involves spontaneous formation of Schiff base bonds between aldehydes (on the support) and amines (on the ligand) and their subsequent stabilization by incubation with a mild reductant (sodium cyanoborohydride). The entire coupling reaction, called reductive amination, occurs in 4 to 6 hours in simple non-amine buffers such as PBS. Coupling efficiency with antibodies and typical proteins is generally greater than 85%, resulting in 1 to 20 mg of immobilized protein per milliliter of agarose resin.
Coupling efficiencies with AminoLink Plus Coupling Resin exceed those obtained with cyanogen bromide (CNBr) activated supports. Furthermore, AminoLink Plus Chemistry forms a bond with the amine-containing ligand that is more stable than with the CNBr method and is uncharged. These properties provide a more leak-resistant immobilization and lower nonspecific binding when used for affinity purification procedures.