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AminoLink Plus Coupling Resin and Kits


A simple method to make affinity purification resin with antibodies or other proteins.

AminoLink Plus Coupling Resin and Immobilization Kits

Thermo Scientific AminoLink Plus Coupling Resin and Kits use aldehyde-activated agarose beads for high-yield covalent coupling of antibodies (proteins) via primary amines to prepare columns for affinity purification.

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.

Highlights:

  • 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
  • Convenient kits and product sizes –choose 1- or 5-column kit containing complete sets of buffers, reagents and versatile spin/drip columns, or select bulk resin.

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 20mg of immobilized protein per milliliter of agarose resin.

AminoLink Plus Resin immobilization chemistry.
Immobilization chemistry of Thermo Scientific AminoLink Plus Coupling Resin. The resin is aldehyde-activated beaded agarose to which antibodies, other proteins and molecules can be covalently attached through primary amines. The result is a reusable affinity resin for use in a variety of purification methods. Learn more about amine-reactive immobilization supports.

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.

High yield protein coupling. Immobilization efficiencies for various proteins on 1mL of AminoLink Plus Resin when immobilized using the pH 7.2 coupling procedure.
Protein Protein (mg)
Reacted
Protein (mg)
Coupled
Coupling
Efficiency (%)
Protein G 4.6 4.0 83
Mouse IgG 4.7 4.5 96
Rat IgG 4.7 4.4 93
GAM-IgG† 0.9 0.8 84
Human IgG 4.8 4.6 97
Human IgM 0.9 0.8 93
† GAM-IgG is goat anti-mouse IgG, a secondary antibody.

References:

  1. Beall, A., et al. (1999). J. Biol. Chem. 274:11344-51.
  2. Nakasato, Y.R., et al. (1999). Clin. Chem. 45:2150-7.
  3. Allan, B.B., et al. (2000). Science 289:444-8.
  4. Lu, R., et al. (2000). J. Neurochem. 74:320-6.

Related Resources:

Review of affinity purification
Review of covalent immobilization methods

Related Products:

Microscale Protein Coupling Kit – uses AminoLink Plus Resin
Pierce Direct Immunoprecipitation Kit – uses AminoLink Plus Resin
AminoLink Reductant (cyanoborohydride)
All Affinity Supports for Ligand Immobilization


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
44894 AminoLink Plus Immobilization Kit, 2mL
Formulation: Multi-component kit
Sufficient For: Immobilizing 2 to 40mg of protein per column

Kit Contents:
AminoLink Plus Columns, 2mL, 5 columns
PBS Pack (makes 500mL), 1 pack
Citrate-Carbonate Buffer Pack (makes 500mL), 1 pack
Sodium Cyanoborohydride Solution, 5M, 0.5mL
Quenching Buffer, 60mL
Wash Solution, 240mL
Column Accessories
5-column kit Product Instructions for product #44894 AminoLink Plus Immobilization Kit, 2mL MSDS for product #44894 AminoLink Plus Immobilization Kit, 2mL Certificate of Analysis for product #44894 AminoLink Plus Immobilization Kit, 2mL $287.00

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20394 AminoLink Plus Immobilization Trial Kit, 2mL
Formulation: Multi-component kit
Sufficient For: Immobilizing 2 to 40mg of protein per column

Kit Contents:
AminoLink Plus Column, 2mL, 1 column
PBS Pack (makes 500mL), 1 pack
Citrate-Carbonate Buffer Pack (makes 500mL), 1 pack
Sodium Cyanoborohydride Solution, 5M, 0.5mL
Quenching Buffer, 15mL
Wash Solution, 50mL
Column Accessories
1-column kit Product Instructions for product #20394 AminoLink Plus Immobilization Trial Kit, 2mL MSDS for product #20394 AminoLink Plus Immobilization Trial Kit, 2mL Certificate of Analysis for product #20394 AminoLink Plus Immobilization Trial Kit, 2mL $105.00

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20501 AminoLink Plus Coupling Resin
Formulation: Aldehyde-activated 4% beaded agarose, slurried in water with sodium azide
Sufficient For: Immobilizing 1 to 20mg of antibody per mL of resin
10mL Product Instructions for product #20501 AminoLink Plus Coupling Resin MSDS for product #20501 AminoLink Plus Coupling Resin Certificate of Analysis for product #20501 AminoLink Plus Coupling Resin $117.00

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20501B AminoLink Plus Coupling Resin
Formulation: Aldehyde-activated 4% beaded agarose, slurried in water with sodium azide
Sufficient For: Immobilizing 1 to 20mg of antibody per mL of resin
Custom MSDS for product #20501B AminoLink Plus Coupling Resin Certificate of Analysis for product #20501B AminoLink Plus Coupling Resin
20505 AminoLink Plus Coupling Resin
Formulation: Aldehyde-activated 4% beaded agarose, slurried in water with sodium azide
Sufficient For: Immobilizing 1 to 20mg of antibody per mL of resin
50mL Product Instructions for product #20505 AminoLink Plus Coupling Resin MSDS for product #20505 AminoLink Plus Coupling Resin Certificate of Analysis for product #20505 AminoLink Plus Coupling Resin $399.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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