Thermo Scientific AminoLink Coupling Resin is crosslinked 4% beaded agarose that has been activated with aldehyde groups to enable covalent immobilization of antibodies and other proteins through primary amines.
Proteins and other molecules with primary amines can be covalently attached (immobilized) to AminoLink Resin to make chromatography columns for use in affinity purification. The aldehyde groups form stable secondary amine bonds with primary amines such as exist in the side chain of lysine (K) residues, which are generally abundant and readily accessible in proteins. Once a protein is immobilized, the prepared affinity resin can be used for a variety of batch and column affinity purification methods involving binding interactions with the immobilized protein. The resin and linkage are stable in most binding and elution conditions typically used in affinity chromatography, enabling prepared resin to be used for multiple rounds of affinity purification procedures.
- AminoLink Coupling Resin – 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)
- Stable, permanent immobilization – Coupling reaction results 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
The AminoLink Resin immobilization reaction involves spontaneous formation of Schiff base bonds between aldehydes and amines and their subsequent stabilization by incubation with a mild reductant. 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 resin.
|Immobilization chemistry of Thermo Scientific AminoLink 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.
Protein coupling efficiency vs. pH of reaction. This experiment involved reaction of 9.58mg of Human IgG with 2mL of Thermo Scientific AminoLink Resin using the standard protocol.
|† Schiff-base formation occurs readily at high pH, but reduction to stable secondary amine bond requires subsequent incubation with sodium cyanoborohydride (AminoLink Reductant) at pH 7.2. The AminoLink Plus Immobilization Kit uses a form of agarose beads with improved flow characteristics and a more convenient protocol for two-step, high-yield coupling.
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Review of affinity purification
Review of covalent immobilization methods
AminoLink Plus Resin and Immobilization Kits – advanced version of AminoLink Resin
Pierce Direct Immunoprecipitation Kit – IP kit based on the AminoLink Method
All Affinity Supports for Ligand Immobilization