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Amine-reactive biotinylation reagent, mid-length
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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.


Bioconjugate Techniques , 2nd ed. By Greg T.Hermanson  (Pierce Biotechnology, Thermo Fisher Scientific, Rockford, IL).  Academic Press  (an imprint of Elsevier):  London, Amsterdam, Burlington, San Diego . 2008. ISBN 978-0-12-370501-3.


• Protein labeling—biotinylate antibodies to facilitate immobilization, purification or detection
• Cell surface labeling—do not penetrate the plasma membrane, biotinylates only surface proteins of whole cells
• Amine-reactive—reacts with primary amines (-NH2), such as lysine side-chains, or the N-terminal-amine
• Cleavable—disulfide bond in spacer arm allows the biotin label to be removed using reducing agents such as DTT
• Soluble—charged sulfo-NHS group increases reagent water solubility compared to ordinary NHS-ester compounds
• Medium length—spacer arm is 24.3 angstroms; it consists of the native biotin valeric acid group extended by a 7-atom chain

Product Citation

Chemical Properties

StorageStore at -20°C
The product is not stable in solution, please dissolve it immediately before use.
Cas No.325143-98-4
SynonymsBiotin disulfide N-hydroxysulfosuccinimide ester
Solubility≥30.335mg/mL in DMSO
Chemical Namesodium;1-[3-[2-[5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoylamino]ethyldisulfanyl]propanoyloxy]-2,5-dioxopyrrolidine-3-sulfonate
SDFDownload SDF
Canonical SMILESC1C(C(=O)N(C1=O)OC(=O)CCSSCCNC(=O)CCCCC2C3C(CS2)NC(=O)N3)S(=O)(=O)[O-].[Na+]
Shipping ConditionEvaluation sample solution : ship with blue ice.All other available size: ship with RT , or blue ice upon request
General tipsFor obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.


Biotinylation method [1]:


468/EV cells and 468/uPAR cells.

Preparation method

Soluble in water, DMSO or DMF.

Reaction Conditions

1 mg/ml, 15minutes on ice


Cells in monolayer culture (1.5 × 106) were washed three times with ice-cold PBS and then treated with sulfo-NHS-SS-biotin (1mg/mL) for 15 minutes on ice. Biotinylation reactions were terminated with 100 mmol/L glycine in PBS. After washing with PBS, cell extracts were prepared in radioimmunoprecipitation assay (RIPA) buffer (20 mmol/L sodium phosphate, 150 mmol/L NaCl (pH 7.4), 1% NP40, 0.1% SDS, and 0.5% deoxycholic acid) with protease inhibitor cocktail. Biotinylated membrane proteins were precipitated with streptavidin-sepharose. Proteins were eluted with SDS sample buffer, resolved by SDS-PAGE, electrotransferred to polyvinylidene difluoride (PVDF) membranes, and probed with primary antibodies.


[1]. Minji Jo, Boryana M. Eastman, Drue L. Webb, et al. Cell Signaling by Urokinase-type Plasminogen Activator Receptor Induces Stem Cell-like Properties in Breast Cancer cells . Cancer Res, 2010;70:8948-8958

Biological Activity

Description Sulfo-NHS-SS-Biotin is a water-soluble, NHS-ester biotinylation reagent

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