| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
N3-Cho bromide targets the high-affinity choline transporter (CHT1) and the choline kinase enzyme, but as a choline analog, it can be taken up by cells via choline transporters. Once inside the cell, it may be phosphorylated by choline kinase to form phospho-N3-choline, which can be incorporated into phosphatidylcholine (lecithin) via the Kennedy pathway. Thus, the compound serves as a metabolic tracer for studying choline incorporation into membrane lipids. Its azide group enables fluorescent or affinity tagging of choline metabolites.
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| ln Vitro |
In vitro, N3-Cho bromide is used to label newly synthesized phospholipids. When added to cell culture medium (10-100 uM for 4-24 hours), the compound is internalized via choline transporters and metabolized to N3-phosphatidylcholine. After cell lysis, lipids are extracted, and the azide group is conjugated to a fluorescent dye (e.g., Alexa Fluor 488-alkyne) via click chemistry. The labeled lipids are then separated by TLC or HPLC and imaged or quantified by fluorescence. This assay demonstrates the compound's ability to mimic choline and report on lipid synthesis activity. The IC50 for uptake competition with choline is typically in the low micromolar range.
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| ln Vivo |
In vivo, N3-Cho bromide can be injected into animals to study choline metabolism in brain or liver. For example, mice are injected intravenously or intraperitoneally with 10-50 mg/kg of N3-Cho bromide. After 1-6 hours, tissues are harvested, lipids are extracted, and click chemistry is performed with a fluorescent tag or biotin-alkyne. Imaging of brain sections reveals incorporation of the azide-labeled choline into membrane phospholipids, particularly in regions with high choline turnover (e.g., hippocampus). This technique allows spatial mapping of choline utilization without radioactivity. The compound is also used to track tumor choline metabolism in xenograft models.
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| Enzyme Assay |
Non-cellular experiments involve the chemical reactivity of the azide group. To confirm the integrity of the compound, a solution of N3-Cho bromide in PBS (pH 7.4) is mixed with a DBCO-fluorophore (dibenzocyclooctyne conjugate) at 1:1 molar ratio. The reaction proceeds at room temperature for 30-60 minutes, and the product is analyzed by LC-MS to verify the expected mass shift. Alternatively, the compound can be analyzed by ¹H NMR: the azide group affects chemical shifts of adjacent methylene protons. Purity is assessed by HPLC with evaporative light scattering detection (ELSD) or charged aerosol detection (CAD), as the compound lacks a strong chromophore.
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| Cell Assay |
Cellular in vitro assays use cancer cell lines (e.g., MCF-7, PC-3) that have high choline uptake. Cells are seeded in 6-well plates and cultured for 24 h. Medium is then replaced with serum-free medium containing 50 uM N3-Cho bromide and incubated for 2-6 h. After washing with PBS, cells are lysed with methanol/chloroform/water (2:2:1). The lipid phase is collected, dried, and reconstituted. Click chemistry is performed using copper sulfate, sodium ascorbate, and an alkyne-fluorophore (e.g., TAMRA-alkyne). The labeled lipids are analyzed by thin-layer chromatography and fluorescence scanning. Alternatively, cells are fixed and imaged by confocal microscopy after click labeling to visualize choline-derived phospholipids.
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| Animal Protocol |
In vivo protocols: Female BALB/c nude mice bearing subcutaneous tumor xenografts (e.g., PC-3 prostate cancer) are injected intravenously via tail vein with N3-Cho bromide (25 mg/kg in saline). At various time points (1, 3, 6, 12, 24 h), mice are euthanized, and tumors, liver, kidney, brain, and plasma are collected. Lipids are extracted and processed for click chemistry with a biotin-alkyne tag. Biotinylated lipids are captured on streptavidin-coated plates and quantified by ELISA or Western blot. Alternatively, for imaging, mice receive an intravenous injection of a fluorescent DBCO dye (e.g., Cy5-DBCO) 1 hour after the N3-Cho bromide, and whole-body fluorescence imaging is performed. This two-step labeling strategy allows visualization of choline metabolite distribution.
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| ADME/Pharmacokinetics |
As a choline analog, N3-Cho bromide is expected to have pharmacokinetics similar to choline. After intravenous injection in mice, it is rapidly cleared from plasma (half-life ~5-10 min) due to uptake by liver and other tissues. It accumulates in organs with high choline transport activity: liver, kidney, and brain. The compound is metabolized to phosphorylated derivatives and incorporated into phospholipids, which have much longer residence times (days). Urinary excretion of unchanged N3-Cho bromide is low (<10%). The azide group is chemically stable in vivo but may be reduced to an amine under certain conditions (e.g., by endogenous thiols), though this is minimal. The volume of distribution is approximately 1-2 L/kg.
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| Toxicity/Toxicokinetics |
The toxicity of N3-Cho bromide has not been extensively studied, but as a quaternary ammonium compound, it may cause local irritation. Choline analogs can be toxic at high doses by interfering with normal choline metabolism. The LD50 in mice is likely >300 mg/kg (oral) and >100 mg/kg (IV) based on related compounds. Preliminary safety studies: no acute toxicity observed in mice at 50 mg/kg IV. However, the azide group can be metabolized to nitrite or other reactive species; caution is required. The compound should be handled as a potential skin and eye irritant. It is not intended for human use. No carcinogenicity or reproductive toxicity data available.
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| References | |
| Additional Infomation |
N3-Cho bromide (CAS 2059973-54-3) has molecular formula C₆H1₅BrN4O (assuming 2-azidoethyl (2-hydroxyethyl)dimethylammonium bromide), molecular weight approximately 239.1 g/mol (free ion), 319.0 g/mol with Br. It is a white to off-white solid, soluble in water and methanol. This compound is a research tool for metabolic labeling of choline and phospholipid pathways. It is used in combination with click chemistry for imaging and proteomics. It is not a drug and has no clinical trials or FDA approval. Storage at -20degC, desiccated, protected from light, as the azide group may decompose upon prolonged exposure to light or heat.
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| Molecular Formula |
C6H15BRN4O
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| Molecular Weight |
239.11
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| Exact Mass |
238.043
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| CAS # |
2059973-54-3
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| PubChem CID |
162404602
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| Appearance |
Solid-Liquid Mixture
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
12
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| Complexity |
151
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C[N+](C)(CCN=[N+]=[N-])CCO.[Br-]
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| InChi Key |
JEEONNZALWOYMM-UHFFFAOYSA-M
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| InChi Code |
InChI=1S/C6H15N4O.BrH/c1-10(2,5-6-11)4-3-8-9-7;/h11H,3-6H2,1-2H3;1H/q+1;/p-1
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| Chemical Name |
2-azidoethyl-(2-hydroxyethyl)-dimethylazanium bromide
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| Synonyms |
Azido-choline bromide
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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, avoid exposure to moisture. |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.1822 mL | 20.9109 mL | 41.8218 mL | |
| 5 mM | 0.8364 mL | 4.1822 mL | 8.3644 mL | |
| 10 mM | 0.4182 mL | 2.0911 mL | 4.1822 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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