AminoLink™ Plus Coupling Resin, 10 mL - FAQs

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.

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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.

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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.

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

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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.

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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).

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Protein samples can be quantified using Thermo Scientific BCA Protein Assay (Cat. No. 23225) or Coomassie Plus Protein Assay (Cat. No. 23236).

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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.

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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.

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No, the gel is stored in sodium azide with no loss in activity. Azide is not a primary amine and is rather unreactive.

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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.

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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.

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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.

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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.

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