| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
Sulfo-NHS-SS-Biotin sodium targets primary amines on proteins and peptides. The sulfo-NHS ester reacts efficiently with primary amines at pH 7-9 to form stable amide bonds. The biotin moiety provides a high-affinity handle for binding to avidin or streptavidin, enabling the detection, purification, or immobilization of labeled proteins. The negatively charged sulfo group renders the compound water-soluble and cell-impermeable, ensuring that it only labels proteins on the cell surface. The cleavable disulfide bond allows for the release of the labeled protein under reducing conditions.
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| ln Vitro |
Sulfo-NHS-SS-biotin is a reagent for marking cell surfaces. The reagent has a negative charge and is incapable of penetrating the cell membrane [1]. Primary amines (NH2), such as the amino termini of polypeptides or lysine side chains, react with sulfo-NHS-SS-biotin [1]. The sulfonic acid group on sulfo-NHS-SS-Biotin keeps it from passing through cell membranes. The release of biotinylated proteins from streptavidin affinity columns is made possible by the cleavable spacer arm of the Thr group [1]. The monolayer grown cells were rinsed with ice-cold PBS before being treated with 1 mg/ml of sulfo-NHS-SS-biotin for 15 minutes. 100 mM glycine in PBS was used to stop the biotinylation processes. Cell extracts were made in RIPA buffer with a protease inhibitor cocktail after being cleaned with PBS. Proteins on biotinylated membranes are precipitated using streptavidin-agarose and then eluted using SDS sample buffer [2].
In vitro, Sulfo-NHS-SS-Biotin sodium is used to label cell-surface proteins for various applications, including protein purification, identification, and quantification. The compound is added to cells or membrane preparations, and the reaction is quenched with a Tris buffer. The labeled proteins are then isolated using avidin or streptavidin affinity chromatography. The disulfide bond allows for the elution of the labeled proteins under reducing conditions with dithiothreitol (DTT) or β-mercaptoethanol. This method is widely used for the identification of cell-surface proteins by mass spectrometry. |
| ln Vivo |
In vivo, Sulfo-NHS-SS-Biotin sodium is not typically used due to its cell-impermeability and rapid hydrolysis in biological fluids. However, it can be used to label cell-surface proteins in vivo by direct injection into tissues, followed by isolation of the labeled proteins. The compound's water solubility and cleavable linker make it a valuable tool for studying protein-protein interactions and cell-surface proteomics.
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| Enzyme Assay |
In vitro enzyme/receptor binding assays are not applicable to Sulfo-NHS-SS-Biotin sodium, as it is a biotinylation reagent rather than a pharmacologically active compound. However, its labeling efficiency can be assessed by Western blot or ELISA using streptavidin-HRP. The extent of biotinylation is determined by comparing the signal intensity of labeled proteins with a biotinylated standard. The cleavability of the disulfide bond is confirmed by treating the labeled proteins with a reducing agent and observing the loss of biotin signal.
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| Cell Assay |
In vitro cellular experiments for Sulfo-NHS-SS-Biotin sodium involve labeling cell-surface proteins. Cells are incubated with the compound in PBS at 4°C for 30 minutes to 1 hour to label surface proteins. The reaction is quenched with a Tris buffer, and the cells are washed. The labeled proteins are then isolated using avidin or streptavidin beads and analyzed by SDS-PAGE and Western blot. The specificity of surface labeling is confirmed by the absence of labeling of intracellular proteins.
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| Animal Protocol |
In vivo animal studies for Sulfo-NHS-SS-Biotin sodium are not typically conducted due to its use as a labeling reagent rather than a therapeutic agent. However, it can be used to label cell-surface proteins in tissues in vivo. The compound is injected directly into the tissue or administered intravenously, and the labeled proteins are isolated from tissue homogenates using avidin or streptavidin affinity chromatography.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties are not applicable to Sulfo-NHS-SS-Biotin sodium, as it is a labeling reagent rather than a therapeutic agent. The compound is rapidly hydrolyzed in aqueous solutions and has a short half-life in biological fluids. Its cell-impermeability limits its distribution to extracellular spaces.
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| Toxicity/Toxicokinetics |
Toxicology is not applicable to Sulfo-NHS-SS-Biotin sodium, as it is a labeling reagent rather than a therapeutic agent. The compound is generally considered safe for handling in the laboratory under standard safety precautions. It is not known to be toxic, mutagenic, or carcinogenic.
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| References |
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| Additional Infomation |
Sulfo-NHS-SS-Biotin sodium is a water-soluble, cell-impermeable, cleavable biotinylation reagent used for the labeling and purification of cell-surface proteins. It reacts with primary amines on proteins to form stable amide bonds. The biotin moiety enables the detection and purification of labeled proteins using avidin or streptavidin. The cleavable disulfide bond allows for the release of labeled proteins under reducing conditions. This compound is a valuable tool for studying cell-surface proteomics, protein-protein interactions, and receptor biology.
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| Molecular Formula |
C₁₉H₂₇N₄NAO₉S₄
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|---|---|
| Molecular Weight |
606.69
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| Exact Mass |
606.056
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| CAS # |
325143-98-4
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| PubChem CID |
71571496
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| Appearance |
White to yellow solid powder
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| LogP |
1.773
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
12
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| Rotatable Bond Count |
15
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| Heavy Atom Count |
37
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| Complexity |
981
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| Defined Atom Stereocenter Count |
3
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| SMILES |
C1[C@H]2[C@@H]([C@@H](S1)CCCCC(=O)NCCSSCCC(=O)ON3C(=O)CC(C3=O)S(=O)(=O)[O-])NC(=O)N2.[Na+]
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| InChi Key |
IBKZNJXGCYVTBZ-IDBHZBAZSA-M
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| InChi Code |
InChI=1S/C19H28N4O9S4.Na/c24-14(4-2-1-3-12-17-11(10-33-12)21-19(28)22-17)20-6-8-35-34-7-5-16(26)32-23-15(25)9-13(18(23)27)36(29,30)31;/h11-13,17H,1-10H2,(H,20,24)(H2,21,22,28)(H,29,30,31);/q;+1/p-1/t11-,12-,13?,17-;/m0./s1
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| Chemical Name |
sodium;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
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| Synonyms |
SulfoNHSSSBiotin sodium; Sulfo NHS SS Biotin sodium
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~125 mg/mL (~206.04 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (3.43 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (3.43 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (3.43 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.6483 mL | 8.2414 mL | 16.4829 mL | |
| 5 mM | 0.3297 mL | 1.6483 mL | 3.2966 mL | |
| 10 mM | 0.1648 mL | 0.8241 mL | 1.6483 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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