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.