| Size | Price | Stock | Qty |
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| 100mg |
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| 250mg |
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| 500mg |
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| Other Sizes |
| Targets |
Biotin NHS targets primary amines (e.g., lysine ε-amino groups and protein N-termini) on proteins, peptides, antibodies, and oligonucleotides. It does not have a biological target in the traditional sense but serves as a chemical labeling reagent. The N-hydroxysuccinimide (NHS) ester reacts with nucleophilic amines to form stable amide bonds. This enables the attachment of biotin to biomolecules for subsequent detection or purification using streptavidin or avidin.
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| ln Vitro |
Advice (This is our suggested protocol, which should be adjusted to suit your particular circumstances as it simply offers guidance). Calculation: Use ≥ 12-fold molar excess of biotin for protein solutions containing 10 mg/mL and ≥ 20-fold molar excess for protein solutions containing 2 mg/mL for optimal outcomes. 1. Determine how many millimoles of the biotin reagent were added to the reaction. mL protein × (mg protein/mL protein) × (mmoL protein/mg protein) × (20 mmoL protein/mmoL protein) = mmoL biotin is the molar excess, which is 20 times. The recommended molar excess of biotin in a protein sample 2 is * 20 = 2 mg/ml. Determine the amount of 10 mM biotin reagent solution (made in step B.3) in microliters that should be added to the reaction using the following formula: mmoL biotin × (1000000 µL/L) × (L/ 10 mmoL) = µL biotin Biotin labeling reaction: 1. 1–10 mg of protein should be dissolved in 0.5–2.0 mL of PBS using the computation in Calculation. 2. Make a 10 mM biotin reagent solution in an organic solvent (DMSO) before using. 3. To the protein solution, add the appropriate volume (μL) of the 10 mM biotin reagent solution. 4. The reaction can be incubated for two hours on ice or for thirty minutes at room temperature. 5. Use Western blotting or ELISA to find tagged proteins.
Biotin NHS does not exhibit pharmacological activity. Its utility is in biotinylation of biomolecules for research applications. It is used to label proteins, antibodies, peptides, and nucleic acids for detection, purification, and immobilization purposes. The biotin-streptavidin interaction (one of the strongest non-covalent interactions) enables highly sensitive detection in ELISA, Western blotting, and immunohistochemistry. Biotin NHS can also be used to prepare biotinylated surfaces or peptides. |
| ln Vivo |
Biotin NHS is not a pharmacologically active compound and does not have direct in vivo activity. Its in vivo behavior is determined by the biotinylated biomolecule to which it is conjugated. Biotinylated antibodies or proteins can be used for in vivo imaging or targeting studies. The compound itself is used as a labeling reagent rather than a therapeutic agent. In vivo studies are conducted on the biotinylated conjugate.
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| Enzyme Assay |
In vitro biotinylation protocols for Biotin NHS typically involve dissolving the compound in anhydrous DMSO or DMF and adding it to the target protein or peptide solution in PBS or HEPES buffer (pH 7.2-8.5). The reaction is incubated at room temperature for 30-120 minutes or at 4°C overnight. The molar ratio of Biotin NHS to target molecule is typically 5-20:1. Unreacted biotin is removed by dialysis, size exclusion chromatography, or desalting columns. The degree of biotinylation is assessed by MALDI-TOF, HABA assay, or streptavidin-based detection. The compound is moisture-sensitive and should be used immediately after preparation.
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| Cell Assay |
For cell-based studies, Biotin NHS is used to label cell surface proteins or to prepare biotinylated ligands for receptor studies. Cells are washed with PBS and incubated with Biotin NHS (typically 0.1-1 mM) in PBS at 4°C for 30 minutes to label surface proteins. After washing, cells are lysed and biotinylated proteins are captured on streptavidin beads for proteomic analysis. Alternatively, biotinylated antibodies or ligands are used to treat cells for receptor binding or uptake studies. The compound itself is not tested for cellular activity.
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| Animal Protocol |
In vivo animal experiments using Biotin NHS typically involve administration of biotinylated antibodies or proteins for imaging or targeting studies. The biotinylated conjugate is administered via IV, IP, or SC routes. Biodistribution is assessed by collecting tissues and detecting biotinylated conjugate using streptavidin-HRP or fluorescent streptavidin. Alternatively, Biotin NHS may be used to label cell surface proteins in vivo for tracking studies. The compound itself is not administered as a therapeutic. All procedures follow institutional animal care guidelines.
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| ADME/Pharmacokinetics |
Biotin NHS (MW 341.38) is moisture-sensitive and should be stored at -20°C in a sealed container away from moisture and light. The compound is unstable in solution and should be prepared fresh immediately before use. It is soluble in DMSO and DMF but hydrolyzes in aqueous solution. The NHS ester reacts rapidly with primary amines. Pharmacokinetic properties are determined by the biotinylated conjugate rather than the compound itself. The compound is for research use only.
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| Toxicity/Toxicokinetics |
Toxicology data for Biotin NHS are limited, as the compound is a research labeling reagent rather than a therapeutic agent. Biotin and NHS ester are generally considered low in toxicity. The compound may cause irritation upon contact. Standard laboratory safety precautions should be followed. The compound is for research use only and not intended for human therapeutic applications. The safety profile of biotinylated conjugates depends on the biomolecule being labeled.
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| References | |
| Additional Infomation |
Biotin NHS (NHS-Biotin) is a research-grade biotinylation reagent for protein, peptide, antibody, and oligonucleotide labeling. Its primary applications include protein detection, purification, and immobilization using streptavidin-biotin technology. The compound is not a drug and has no clinical applications. It is widely used in molecular biology, biochemistry, and immunology research. The compound is commercially available from various chemical suppliers for research purposes only.
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| Molecular Formula |
C14H19N3O5S
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|---|---|
| Molecular Weight |
341.3828
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| Exact Mass |
341.104
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| CAS # |
35013-72-0
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| PubChem CID |
6710714
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| Appearance |
White to off-white solid powder
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| Density |
1.5±0.1 g/cm3
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| Melting Point |
212-214 °C
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| Index of Refraction |
1.616
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| LogP |
-1.41
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
23
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| Complexity |
525
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| Defined Atom Stereocenter Count |
3
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| SMILES |
S1C([H])([H])[C@@]2([H])[C@@]([H])([C@]1([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C(=O)ON1C(C([H])([H])C([H])([H])C1=O)=O)N([H])C(N2[H])=O
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| InChi Key |
YMXHPSHLTSZXKH-RVBZMBCESA-N
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| InChi Code |
InChI=1S/C14H19N3O5S/c18-10-5-6-11(19)17(10)22-12(20)4-2-1-3-9-13-8(7-23-9)15-14(21)16-13/h8-9,13H,1-7H2,(H2,15,16,21)/t8-,9-,13-/m0/s1
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| Chemical Name |
(2,5-dioxopyrrolidin-1-yl) 5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoate
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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: (1). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. (2). This product is not stable in solution, please use freshly prepared working solution for optimal results. |
| 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 : ~62.5 mg/mL (~183.08 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (6.09 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 (6.09 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 (6.09 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 | 2.9293 mL | 14.6464 mL | 29.2929 mL | |
| 5 mM | 0.5859 mL | 2.9293 mL | 5.8586 mL | |
| 10 mM | 0.2929 mL | 1.4646 mL | 2.9293 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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