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Thermo Scientific™ EZ-Link™ Sulfo-NHS-LC-Biotin
Description
Thermo Scientific™ EZ-Link Sulfo-NHS-LC-Biotin is an intermediate-length, water-soluble biotinylation reagent for labeling antibodies, proteins and other molecules that have primary amines.
- Protein labeling - biotinylate antibodies to facilitate immobilization, purification or detection using streptavidin resins or probes
- Cell surface labeling - biotinylates only surface proteins of whole cells because the negatively charged reagent does not permeate cell membranes
- Amine-reactive - reacts with primary amines (-NH2), such as lysine side-chains, or the amino-termini of polypeptides
- Soluble - charged sulfo-NHS group increases reagent water solubility compared to ordinary NHS-ester compounds
- Irreversible - forms permanent amide bonds; spacer arm cannot be cleaved
- Medium length - spacer arm (total length added to target) is 22.4 angstroms; provides excellent balance between adequate length (minimal steric hindrance for biotin binding) and modest mass
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Specifications
Specifications
| Description | EZ-Link Sulfo-NHS-LC-Biotin |
| Product Type | Sulfo-NHS-LC-Biotin |
| Quantity | 100 mg |
| Content And Storage | Store desiccated at -20°C. Shipped at ambient temperature. |
| Product Line | EZ-Link |
| Cell Permeability | Cell-Impermeant |
| Chemical Reactivity | Amine |
| Reactive Moiety | Active Ester, Succinimidyl Ester, Sulfo-NHS Ester |
| Solubility | DMF (Dimethylformamide), DMSO (Dimethylsulfoxide), Water |
| Label or Dye | Biotin |
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Frequently Asked Questions (FAQs)
No. NHS esters hydrolyze very quickly. The container must be brought to room temperature (RT) prior to opening, then the amount of biotinylation reagent weighed, dissolved and used immediately.
Yes. Because this molecule dissolves readily in polar solutions and is itself charged by the sodium sulfoxide group on the succinimidyl ring, it cannot permeate the cell wall or membrane of an intact cell. As long as the cell is intact, only the primary amines on the surface will be biotinylated.
Tris is a primary amine, which will directly compete with protein primary amines for reaction with the NHS-ester activated biotin reagent. However, once a protein is labeled and desalted, Tris buffer can be used for storage or assay buffers, as appropriate for the protein and application.
The molar excess to use depends on the concentration of the ligand to be biotinylated. Although the literature contains references that use between 2-50 molar excess of biotin, we recommend the following: 10-fold molar excess for 10 mg/mL solution of protein; 25-fold molar excess for 2 mg/mL solution of protein.
No, the EZ-Link Sulfo-NHS-LC-Biotin, No-Weigh Format (Cat. No. A39257) should not be dissolved in water for storage. We recommended storing EZ-Link Sulfo-NHS-LC-Biotin, No-Weigh Format at -20 degrees C with desiccant as it is very moisture sensitive. Furthermore, to avoid moisture condensation onto the product, equilibrate vial to room temperature before opening.
Safety and Handling
For Research Use Only. Not for use in diagnostic procedures.
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