Sulfo-NHS-SS-Biotin Kit
Sulfo-NHS-SS-Biotin is a water-soluble biotinylation reagent frequently employed in biomedical research to chemically label proteins, antibodies, peptides, or other amine-containing biomolecules. Its labeling mechanism is based on the reaction between the reagent's sulfo-N-hydroxysuccinimide (Sulfo-NHS) ester group and primary amines (-NH2) of target molecules, forming stable amide linkages. The incorporated disulfide bond (-SS-) within the reagent structure enables subsequent cleavage under reducing conditions, allowing reversible attachment and efficient molecule recovery. Common research applications include purification, immobilization, detection, and affinity-based analyses, such as protein interaction studies, western blotting, immunoprecipitation, and cell-surface biotinylation experiments.
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Related Biological Data
| Sufficient For: 10 labeling reactions, each with 1 to 10 mg of antibody | |||
| Streptavidin | 10 mg | Sulfo-NHS-SS-Biotin | 25 mg |
| HABA Solution | 1 mL | PBS Pack (makes 1 L) | 1 pack |
| Sephadex G-25 in PD-10 Desalting Columns | 8.3 mL, 10 columns | ||
| Store biotin and streptavidin reagents at -20°C. Store remaining kit components at 4°C. | |||
| Label or Dye: | Biotin | Product Size: | 10 reactions |
| Labeling Scale: | 1-10 mg | Labeling Target: | Proteins (General), Antibodies (General) |
• Amine-reaction—Reacting with primary amines (-NH2), such as lysine side-chains, or the amino-termini of polypeptides.
• Labeling antibody—This kit can label antibodies to facilitate immobilization, purification or detection.
• Labeling protein—This kit can label proteins to facilitate immobilization, purification or detection.
• Labeling Cell surface molecule—This kit can label the cell surface proteins because the negatively charged reagent does not permeate cell membranes.
• Reversible—Disulfide bond in spacer arm allows the biotin label to be removed using reducing agents such as DTT; only a small sulfhydryl group remains attached to the molecule.
• Medium length—Spacer arm (total length added to target) is 24.3 angstroms; it consists of the native biotin valeric acid group extended by a 7-atom chain
• Solubility increased—Sulfo-NHS group increases reagent water solubility compared to ordinary NHS-ester compounds.
Sulfo-NHS-SS-biotin (sulfosuccinimidyl-20(biotinamido)ethyl-1,3-dithiopropionate) is a long-chain cleavable amine-reactive biotinylation reagent. The presence of the negatively charged sulfonate group in the chemical structure of sulfo-NHS-SS-biotin makes it a water-soluble biotinylation reagent that can be directly added to aqueous reactions without prior dissolution of organic solvents. Although no prior dissolution is required, an aqueous stock solution of sulfo-NHS-SS-biotin must be prepared rapidly and used immediately in case of the occurrence of hydrolysis of the active ester. Sulfo-NHS-SS-biotin has been used to react with amine-containing proteins and other molecules forming a complex which further interacts with avidin or streptavidin probes and to purify targeted molecules using affinity chromatography on a column of immobilized avidin or streptavidin.
