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CarboxyLink Coupling Resin and Kits


Immobilize peptides via carboxyl groups to create an affinity column.

CarboxyLink Coupling Resin and Kits

Thermo Scientific CarboxyLink Coupling Resin and Kits are 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 (or polyacrylamide UltraLink Support) 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.

Highlights:

  • CarboxyLink Coupling Resin – DADPA-activated crosslinked 4% beaded agarose (or DADPA-activated UltraLink Support, a beaded polyacrylamide resin)
  • Efficient immobilization – Couple 1 to 2mg of peptide per milliliter of resin (CarboxyLink Agarose Resin activated with greater than 16µmol amine milliliter of resin; DADPA on UltraLink Support activated with greater than 40µmol 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
  • Convenient kits and product sizes – choose 5-column kits with complete sets of buffers, crosslinker and versatile spin/drip columns containing either type of resin (agarose or polyacrylamide) or choose stand-alone resin for other uses.

Product Details:

Immobilization chemistry of CarboxyLink Resin
Immobilization chemistry for carboxyl-containing molecules. EDC crosslinker reacts with carboxylates to create an amine-reactive intermediate, resulting in covalent attachment to the DADPA-activated Resin (Thermo Scientific CarboxyLink Coupling Resin). Learn more about carboxyl-reactive immobilization methods and about EDC crosslinking chemistry.

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.

References:

  1. Yoo, B.C., et al. (2002). J. Biol. Chem. 277, 15325-15332.
  2. Shenoude, A., et al. (2002). J. Histochem. Chytochem. 50(7), 885-889.
  3. Morrisette, J.M., et al. (2003). J. Ex. Biol. 206, 805-812.
  4. Xainli, J., et al. (2003). Infect. Immunity 71(5), 2508-2515.
  5. Cawley, N.X., et al. (2003). J. Biol. Chem. 278(8), 5523-5530.

Related Resources:

CarboxyLink Coupling Resins and Kit FAQ
Review of affinity purification
Review of covalent immobilization methods
Carbodiimide (EDC) crosslinker chemistry

Related Products:

EDC Crosslinker
MES Buffered Saline Packs
Other Activated Affinity Resins


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
20266 CarboxyLink Coupling Resin
Formulation: DADPA-activated crosslinked 4% beaded agarose, slurried in water with sodium azide
Sufficient For: Immobilizing 0.5 to 1mg of peptide per mL of resin
25mL Product Instructions for product #20266 CarboxyLink Coupling Resin MSDS for product #20266 CarboxyLink Coupling Resin Certificate of Analysis for product #20266 CarboxyLink Coupling Resin $157.00

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20266B CarboxyLink Coupling Resin
Formulation: DADPA-activated crosslinked 4% beaded agarose, slurried in water with sodium azide
Sufficient For: Immobilizing 0.5 to 1mg of peptide per mL of resin
Custom MSDS for product #20266B CarboxyLink Coupling Resin Certificate of Analysis for product #20266B CarboxyLink Coupling Resin
44899 CarboxyLink Immobilization Kit, 2mL
Formulation: Multi-component kit
Sufficient For: Immobilizing 1 to 2mg of peptide per column

Kit Contents:
CarboxyLink Columns, 2mL, 5 columns
EDC Crosslinker, 5 x 60mg
MES Buffered Saline Pack (makes 500mL), 1 pack
Wash Solution, 120mL
Column Accessories
5-column kit Product Instructions for product #44899 CarboxyLink Immobilization Kit, 2mL MSDS for product #44899 CarboxyLink Immobilization Kit, 2mL Certificate of Analysis for product #44899 CarboxyLink Immobilization Kit, 2mL $423.00

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53154 CarboxyLink Immobilization Kit with UltraLink Support, 2mL
Formulation: Multi-component kit
Sufficient For: Immobilizing 1 to 2mg of peptide per column

Kit Contents:
DADPA UltraLink Columns, 2mL, 5 columns
EDC Crosslinker, 5 x 60mg
MES Buffered Saline Pack (makes 500mL), 1 pack
Wash Solution, 120mL
Column Accessories
5-column kit Product Instructions for product #53154 CarboxyLink Immobilization Kit with UltraLink Support, 2mL MSDS for product #53154 CarboxyLink Immobilization Kit with UltraLink Support, 2mL Certificate of Analysis for product #53154 CarboxyLink Immobilization Kit with UltraLink Support, 2mL $465.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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