CarboxyLink™ Coupling Resin
CarboxyLink™ Coupling Resin
Thermo Scientific™

CarboxyLink™ Coupling Resin

Thermo Scientific CarboxyLink Coupling Resin is for covalent immobilization of peptides or other carboxyl-containing (—COOH) molecules to a porous, beadedRead more
Have Questions?
Catalog number 20266
Price (USD)
256.00
Each
Add to cart
Request bulk or custom format
Price (USD)
256.00
Each
Add to cart
Thermo Scientific CarboxyLink Coupling Resin is for covalent immobilization of peptides or other carboxyl-containing (—COOH) molecules to a porous, beaded resin for use in affinity purification procedures.

CarboxyLink Resin is crosslinked beaded agarose that has been activated with diamino-dipropylamine (DADPA) to contain long spacer arms, each with a primary amine at the end. When incubated with the resin and the carbodiimide crosslinker EDC (included in the CarboxyLink Immobilization Kit), carboxyl-containing molecules become permanently attached to the support by stable amide bonds. Once a molecule is coupled, the prepared affinity column can be used multiple times in typical protein affinity purification procedures. CarboxyLink Coupling Resins can also be used to immobilize other kinds of molecules using alternative amine-reactive crosslinking chemistries.

Features of the CarboxyLink Coupling Resin:

CarboxyLink Coupling Resin—DADPA-activated crosslinked 4% beaded agarose
Efficient immobilization—Couple 1 to 2 mg of peptide per milliliter of resin (CarboxyLink Agarose Resin activated with greater than 16 μMol amine milliliter of resin; DADPA on UltraLink Support activated with greater than 40 μMol amine milliliter of resin)
Stable linkage—immoblization results in covalent attachment of carboxyl groups by amide bonds, allowing for multiple rounds of affinity purification with one batch of prepared resin
Flexible and gentle coupling conditions—immobilization reaction completed in simple MES or other non-amine and non-carboxyl, near-neutral buffer, with or without organic solvent.
Ideal for unmodified peptides—immobilizes peptides with high capacity and various orientations without steric hindrance, allowing for effective use in affinity purification of specific antibodies

Because EDC crosslinks carboxylates to primary amines, peptides and other molecules that contain both of these functional groups will be variously polymerized as well as immobilized to the CarboxyLink Resin. For peptides whose only carboxyl and amino groups are at the termini, coupling with the CarboxyLink Kit will result in end-to-end chains of peptide that are linked by their C-terminal ends to the support. Generally, this structure provides for high capacity and low steric hindrance for use in affinity purification. However, in the case of peptides containing multiple primary amines (lysine) or carboxylates (glutamic and aspartic acid) in the middle of their sequence, the CarboxyLink Coupling Method may cause key binding sites to be blocked, resulting in high coupling capacity but poor binding capacity for affinity purification. Alternative methods for immobilization include AminoLink Plus Coupling Resin, UltraLink Biosupport and SulfoLink Coupling Resin.

Related Products
CarboxyLink™ Immobilization Kit, 2 mL
CarboxyLink™ Immobilization Kit with UltraLink™ Support, 2 mL
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Capacity (Metric)25 mL
DescriptionCarboxyLink Coupling Resin
FormLiquid
Product LineCarboxyLink™
TargetPrimary Amine
TypeResin
Stationary PhaseAmine
FormatBottle
Column TypeAffinity, Agarose Resin
Unit SizeEach
Contents & Storage
Store at 4°C.

Frequently asked questions (FAQs)

What is the difference between the Thermo Scientific CarboLink (now Glycolink) and CarboxyLink Resins?

The CarboxyLink Resin is for immobilizing carboxyl-containing biomolecules after EDC activation. CarboLink Resin is hydrazide-activated and immobilizes glycoproteins. Carbolink has been replaced with glycolink immobilization kits and resins. Glycolink can also be used to immobilize steroids or other molecules containing ketones. Both resins are capable of coupling ligands via carboxylic acids using EDC (Cat. No. 22980). Both immobilization chemistries are available on UltraLink Resin as UltraLink Hydrazide (glycoproteins) and UltraLink DADPA (carboxyl) Resins.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

How many purifications can be performed using the same affinity column with CarboxylLink or DADPA UltraLink resin?

The stability of the immobilized protein and the type of elution buffer used determines how many times a column can be reused. Typically, the columns can be reused at least 10 times without significant loss in purification efficiency.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

My peptide is not water-soluble; can I use other solvents for conjugation when using the CarboxylLink or DADPA UltraLink resin?

Yes. When coupling water-insoluble peptides or other molecules, use water-miscible solvents such as ethanol, methanol, DMSO or DMF. Dissolve the peptide in 100% of the water-miscible solvent first and then add this solution to the Conjugation Buffer. Organic solvent concentrations up to 50% in the coupling reaction are compatible. When using high concentrations of organic solvent (> 25%), gradually equilibrate the resin into the organic solvent. For example, wash the resin with 2-3 column volumes of each of the following solutions before adding the sample in 50% organic solvent:

- 95% aqueous, 5% organic solvent
- 85% aqueous, 15% organic solvent
- 75% aqueous,25% organic solvent
- 60% aqueous, 40% organic solvent
- 50% aqueous, 50% organic solvent
Reverse the percent organic solvent and re-equilibrate the resin in 100% aqueous if purifying a sample in aqueous buffer.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

How do I determine the degree of antibody coupling to the CarboxylLink or DADPA UltraLink resin?

Protein samples can be quantified using Thermo Scientific BCA Protein Assay (Cat. No. 23225) or Coomassie Plus (Bradford) Protein Assay (Cat. No. 23236). Alternatively, coupling can be estimated by measuring the protein concentration in solution before and after coupling. The difference in the two measurements is the amount coupled to the resin. Make sure to factor in any change in sample volume.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Are the CarboxylLink and DADPA UltraLink Resins available as kits?

CarboxyLink Coupling Resin is available as a kit (Cat. No. 44899), which contains sufficient components to prepare five reusable affinity columns. The resin is also available separately (Cat. No. 20266, 25 mL). DADPA UltraLink is available only as a kit (Cat. No. 53154). The kits do not contain buffers for affinity purification.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.