missing translation for 'onlineSavingsMsg'
Learn More
  1. Home
  2. Products
  3. Protein Analysis Reagents
  4. Protein Purification
  5. Protein Purification Reagents and Kits
  6. Thermo Scientific™ AminoLink™ Plus Coupling Resin

Thermo Scientific™ AminoLink™ Plus Coupling Resin

Catalog No. PI20501
Change view
Click to view available options
Quantity:
10 mL
50 mL
2 product options available for selection
Product selection table with 2 available options. Use arrow keys to navigate and Enter or Space to select.
Catalog No. Quantity
PI20501 10 mL
PI20505 50 mL
Use arrow keys to navigate between rows. Press Enter or Space to select a product option. 2 options available.
2 options
Catalog No. PI20501 Supplier Thermo Scientific™ Supplier No. 20501
Only null left

Description

Covalently couple antibodies and other proteins via primary amines (-NH2) for affinity purification with these aldehyde-activated agarose beads.

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

The resins and kits use activated beaded agarose and a robust coupling chemistry to immobilize proteins and other ligands through primary amines (-NH2) to the resin. Once an antibody or other ligand is immobilized, the prepared affinity resin can be used for a variety of 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.

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 beads.

Highlights:

  • AminoLink Plus 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); regular (PBS, pH 7.2) and enhanced (borate, pH 10) coupling protocols provided

  • 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

  • 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; bulk quantities available for manufacturing applications

Includes:

Kits contain AminoLink Plus Columns, neutral and enhanced coupling buffers (Part No. 44894 only), quenching buffer, wash solution, sodium cyanoborohydride solution, and column accessories

Specifications

Formulation Aldehyde-activated 4% beaded agarose, slurried in water with sodium azide
Content And Storage Store at 4°C.
Form Liquid
Product Line AminoLink
Quantity 10 mL
Target Antibody, Proteins through Primary Amine
Type Resin
Stationary Phase Aldehyde
Format Bottle
Column Type Affinity, Agarose Resin

Frequently Asked Questions (FAQs)

What is the binding capacity of the AminoLink Coupling Resin or aminoLink Plus Resin?

AminoLink Coupling Resin can bind approximately 10 mg of IgG/mL of resin bed, while AminoLink Plus Resin can bind up to 15-20 mg of IgG/mL of resin bed. The upper limit for binding peptides is approximately 1 mg/mL of resin for both support types.

What are the differences between the two supports for the AminoLink Plus Resin?

The AminoLink Plus Resin uses a type of agarose that has better flow properties, which may shorten the time needed for drip-column methods. It is also activated at a higher level, allowing greater ligand to be immobilized.
My ligand is not soluble in the coupling buffer when using the AminoLink Coupling Resins or Kits. What can I do?

Coupling reactions with AminoLink Resins are compatible with 3-4 M fresh urea or guanidine. (However, old solutions of urea may contain ammonia, which will interfere with ligand immobilization.) Alternatively, dissolve the peptide in 100% DMSO. Add the peptide in DMSO to coupling buffer so that the DMSO does not exceed 20% of the final solution.

How many purifications can be performed using the same AminoLink or AminoLink Plus Column?

Columns can be reused at least 10 times without loss of activity.

How long will the coupling reaction take when using the AminoLink Resins and Kits?

The immobilizatoin reaction takes six hours; the entire procedure may require seven to eight hours. This time can be decreased if the high pH protocol for AminoLink Plus Resin is used.
How can I perform affinity purifications with conjugated AminoLink and AminoLink Plus Affinity Columns?

Equilibrate the prepared column with three to five bed volumes of an appropriate binding buffer. Add 1 mL of sample for each 2 mL bed-volume (serum should be diluted at least 1:1 with binding buffer). Add an additional 200µL of binding buffer to ensure that the entire sample has entered the gel bed. Cap the column bottom and top. Incubate one hour. Wash away non-bound proteins with five to seven bed-volumes of binding buffer or 1 M NaCl. Elute the bound sample by adding small fractions (0.5-1.0 mL) of elution buffer such as Thermo Scientific IgG Elution Buffer (Cat. No. 21004).
How do I determine the degree of conjugation of the immobilized ligand when using the AminoLink Resins and Kits?

Protein samples can be quantified using Thermo Scientific BCA Protein Assay (Cat. No. 23225) or Coomassie Plus Protein Assay (Cat. No. 23236).
How do AminoLink Resins and Kits work?

Both AminoLink Coupling Resin and AminoLink Plus Coupling Resin consist of a crosslinked beaded agarose that is derivatized to have aldehydes present for coupling. The aldehydes will react with primary amines to form reversible Schiff's bases. After this double bond reacts with sodium cyanoborohydride, the process of reductive amination produces a permanent covalent bond. The aldehyde groups attach directly to the beads without any spacer arms.
Can you provide the shelf-life for AminoLink Plus Coupling Resin?

AminoLink Plus Coupling Resin is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended. Please see section 8.1 of our Terms & Conditions of Sale (https://www.thermofisher.com/content/dam/LifeTech/Documents/PDFs/Terms-and-Conditions-of-Sale.pdf) for more details.
Does sodium azide interfere with AminoLink Support coupling?

No, the gel is stored in sodium azide with no loss in activity. Azide is not a primary amine and is rather unreactive.
How do I know whether to use pH 7.2 or pH 10 for coupling my protein to AminoLink Plus Gel?

Coupling at pH 10 generally gives some increase in efficiency but may damage some ligands and requires a more involved procedure. If stability of the ligand is in question or the increase in coupling efficiency is of minor importance, use the pH 7.2 coupling procedure. This procedure has fewer steps and will still give good coupling efficiency.
What is the difference between AminoLink Coupling Resin and NHS-Activated Agarose?

Both react with primary amines, but AminoLink Coupling Resin is a two-step conjugation. First a Schiff base is formed between the amine and the aldehyde on the resin, which is then reduced to a stable secondary amine with sodium cyanoborohydride, whereas NHS-Activated Agarose utilizes an NHS ester to form an amide bond in a one-step conjugation.
What is the difference between AminoLink and AminoLink Plus supports?

AminoLink Plus Coupling Resin is activated at a higher level and has higher flow rates than the original AminoLink Coupling Resin, resulting in higher capacity and faster purification. AminoLink Supports can be used to immobilize any molecule with a primary amine.
How do your AminoLink Coupling Resin and AminoLink Plus Coupling Resin work?

AminoLink and AminoLink Plus Supports are activated with aldehyde groups which will react with primary amines to form Schiff bases, which are reduced to stable, non-reversible secondary amines. Coupling efficiency often exceeds 85% with this support.

For Research Use Only. Not for use in diagnostic procedures.