SulfoLink™ Coupling Resin, 10 mL - FAQs

View additional product information for SulfoLink™ Coupling Resin - FAQs (20401, 20404, 20402)

10 product FAQs found

What is the binding capacity of SulfoLink Resin?

SulfoLink Resin can bind approximately 5 mg of reduced IgG per mL of support. For peptides, the capacity is approximately 1 mg of peptide per mL of gel. Capacity for other compounds will vary.

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

When using the SulfoLink Coupling Resins and Kits, my peptide is not soluble in the coupling buffer. What can I do?

SulfoLink Immobilization is compatible with 3-4 M fresh urea or guanidine. Alternatively, dissolve the peptide in 100% DMSO. Add the peptide in DMSO to the coupling buffer so that the DMSO does not exceed 20% of the final solution.

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

Must I block the resin before each use when using the SulfoLink Coupling Resins and Kits?

No. The resin will no longer react with subsequent sulfhydryls after the ligand is attached and the remaining active sites are blocked.

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

How many purifications can I perform using the same SulfoLink Column?

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

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How does SulfoLink Immobilization work?

SulfoLink Resin is crosslinked 6% beaded agarose that has been derivatized with a 12-atom spacer arm that ends in an iodoacetyl group. This spacer arm is helpful for peptides or large proteins that might otherwise be sterically hindered in binding their ligand. The iodoacetyl group reacts with free sulfhydryls at pH 7.5-8.5 to form covalent thioether bonds. The sulfhydryl must be present in the reduced form because disulfides will not react with the iodoacetyl group.

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

How do I reduce my sample and confirm the reduction when using the SulfoLink Coupling Resins and Kits?

The kit includes the reductant beta-Mercaptoethylamine (beta-MEA, Cat. No. 20408); however, the following reductants can also be used: dithiothreitol (DTT), Cat. No. 20290; ß-MEA Cat. No. 35600 and 35601; and TCEP-HCl Cat. No. 20490. You can also use Ellman's Reagent (Cat. No. 22582) to confirm the presence of sulfhydryls. Ellman's Reagent reacts with reduced sulfhydryls to produce a distinctive yellow color that is readable at 412 nm. After it is reduced, desalt the sample directly into an appropriate buffer and allow it to react with the column. This will prevent disulfide formation from reforming.

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How do I perform affinity purifications with conjugated SulfoLink Affinity Columns?

Equilibrate the column with 3-5 bed-volumes of an appropriate binding buffer. Add 1 mL of sample for each 2 mL column (serum should be diluted at least 1:1 with binding buffer). Add an additional 200 µl of binding buffer to ensure that the entire sample has entered the gel bed. Cap the column bottom and top. Incubate the column for 1 hour. Wash away non-bound proteins with 5-7 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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I want to purify an antibody against a peptide that has a cysteine moiety. What support should I use?

SulfoLink Coupling Resin or UltraLink Iodoacetyl Resin would be good choices. They utilize the same chemistry to react with the reduced thiol group.

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What is the difference between SulfoLink and CarboxyLink Coupling Resin supports?

SulfoLink Coupling Resin is iodoacetyl-activated agarose that reacts with reduced thiols to form a stable thioether linkage. CarboxyLink Coupling Resin is an amine-derivatized agarose that utilizes the carbodiimide crosslinker, EDC to conjugate the carbon of carboxylic acid to the nitrogen of the immobilized primary amine (hydrazide) to form a permanent amide bond.

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How does the SulfoLink Coupling Resin work?

Iodoacetyl-derivatized support reacts with free sulfhydryl groups on a protein/peptide or other biological molecule, resulting in a stable thioether bond (non-reducible). This support is ideal for peptides with a terminal cysteine or for orienting monoclonal antibodies that have amines in the antigen-binding site.

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