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AminoLink Coupling Resin and Kit


Easily immobilize antibodies and other proteins for use in affinity purification.

AminoLink Coupling Resin

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.

Highlights:

  • 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

Product Details:

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.

AminoLink Resin immobilization chemistry.
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.
Reaction pH Coupling Efficiency
4 91.8%
5 92.7%
6 89.1%
7 87.3%
8 † 85.3%
9 † 94.9%
10 † 98.4%
† 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.

References:

  1. Assad, F.F., et al. (2001). J. Cell Biol. 152:531-43.
  2. Cheadle, C., et al. (1994). J. Biol. Chem. 269:24034-9.
  3. Cofano, F. (1990). J. Biol. Chem. 265:4064-4071.
  4. Czernik, P.J., and Hurlburt, B.K. (1994). J. Biol. Chem. 269:27869-75.
  5. DeSilva, B.S. and Wilson, G.S. (1995). J. Immunol. Meth. 188:9-19.
  6. Essler, M., et al. (2002). Proc. Natl. Acad. Sci. USA. 99:2252-7.
  7. Fries, D.M., et al. (2003). J. Biol. Chem. 278:22901-7.
  8. James, L.G., et al. (1994). J. Biol. Chem. 269:14182-90.
  9. Kierszenbau, A.L., et al. (2003). Mol. Biol. Cell 14:4628-40.
  10. Li, H. and Pajor, A.M. (2003). Am. J. Physiol. Cell Physiol. 285:1188-96.
  11. Rivero-Lezcano, O.M., et al. (1994). J. Biol. Chem. 269:17363-6.
  12. Rossig, L. et al. (2001). Mol. Cell. Biol. 21: 5644-5657.
  13. Zuk, P.A. and Elferink, L.A. (2000). J. Biol. Chem. 275:26754-64.

Related Resources:

Review of affinity purification
Review of covalent immobilization methods

Related Products:

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


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
20381 AminoLink Coupling Resin
Formulation: Aldehyde-activated crosslinked 4% beaded agarose, slurried in water with sodium azide
Sufficient For: Immobilizing 1 to 10mg of antibody per mL of resin
10mL Product Instructions for product #20381 AminoLink Coupling Resin MSDS for product #20381 AminoLink Coupling Resin Certificate of Analysis for product #20381 AminoLink Coupling Resin $115.00

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20382 AminoLink Coupling Resin
Formulation: Aldehyde-activated crosslinked 4% beaded agarose, slurried in water with sodium azide
Sufficient For: Immobilizing 1 to 10mg of antibody per mL of resin
50mL Product Instructions for product #20382 AminoLink Coupling Resin MSDS for product #20382 AminoLink Coupling Resin Certificate of Analysis for product #20382 AminoLink Coupling Resin $375.00

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20382B AminoLink Coupling Resin
Formulation: Aldehyde-activated crosslinked 4% beaded agarose, slurried in water with sodium azide
Sufficient For: Immobilizing 1 to 10mg of antibody per mL of resin
Custom MSDS for product #20382B AminoLink Coupling Resin Certificate of Analysis for product #20382B AminoLink Coupling Resin
44890 AminoLink Immobilization Kit
Formulation: Multi-component kit
Sufficient For: Immobilizing 2 to 20mg of protein per column

Kit Contents:
AminoLink Columns, 2mL, 5 columns
AminoLink Coupling Buffer (pH 7.0), 250mL
Quenching Buffer, 50mL
Wash Buffer, 240mL
Sodium Cyanoborohydride Solution, 5M, 0.5mL
Column Accessories
5-column Kit Product Instructions for product #44890 AminoLink Immobilization Kit MSDS for product #44890 AminoLink Immobilization Kit Certificate of Analysis for product #44890 AminoLink Immobilization Kit $260.00

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44892 AminoLink Reductant
Formulation: Sodium cyanoborohydride (NaCNBH3); white to yellow crystalline powder; MW 62.84; CAS# 25895-60-7
Sufficient For: Preparing 6.36mL of a typical 100X (5M) solution for reductive amination protocols.
2 x 1g Product Instructions for product #44892 AminoLink Reductant MSDS for product #44892 AminoLink Reductant Certificate of Analysis for product #44892 AminoLink Reductant $44.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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