| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
L-Azidohomoalanine does not have a classical pharmacological target. Its biological function is based on its ability to be recognized by methionyl-tRNA synthetase and incorporated into proteins in place of methionine. This makes it a substrate for the cellular protein synthesis machinery rather than a receptor ligand or enzyme inhibitor. The azide group is bioorthogonal, meaning it does not react with endogenous cellular components, allowing selective labeling of newly synthesized proteins. After incorporation, the azide can be reacted with alkyne-functionalized fluorophores, biotin, or other probes for detection, enrichment, or imaging applications.
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|---|---|
| ln Vitro |
One ligand is for an E3 ubiquitin ligase, and the other is for the target protein; these two ligands are joined by a linker to form PROTACs. The intracellular ubiquitin-proteasome system is utilized by PROTACs to specifically destroy target proteins[1].
In vitro activity data for L-Azidohomoalanine relate to its incorporation efficiency and labeling performance. In methionine-auxotrophic cell lines (e.g., HEK293T, HeLa), replacement of methionine with L-Azidohomoalanine in the culture medium results in efficient incorporation into newly synthesized proteins. Typical concentrations of 50-100 microM in methionine-free medium support protein synthesis and labeling. Incorporation efficiency is assessed by click chemistry with alkyne-fluorophore conjugates followed by SDS-PAGE and fluorescence imaging. The compound is non-toxic at working concentrations, with cell viability maintained above 90% in standard assays. |
| ln Vivo |
In vivo activity data for L-Azidohomoalanine are primarily from metabolic labeling studies in animal models. When administered to mice, L-Azidohomoalanine is incorporated into newly synthesized proteins in various tissues. This enables in vivo protein synthesis rate measurement and newly synthesized protein profiling. The compound is typically administered intraperitoneally at doses of 10-50 mg/kg, and tissues are collected after 1-24 hours for click chemistry labeling and analysis. This approach has been used to study protein synthesis in the brain, liver, and other organs. The compound is well tolerated at these doses.
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| Enzyme Assay |
For non-cellular protein labeling assays, cell-free protein synthesis systems or purified protein systems are used. L-Azidohomoalanine is added to in vitro translation reactions at 50-100 microM in methionine-free conditions. After translation, the azide-labeled protein is reacted with a dibenzocyclooctyne (DBCO)-fluorophore conjugate via copper-free click chemistry. The reaction is performed in PBS at room temperature for 1-2 hours. Labeled proteins are analyzed by SDS-PAGE and fluorescence imaging or by mass spectrometry for identification.
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| Cell Assay |
For cell-based protein labeling assays, methionine-auxotrophic cells (HEK293T, HeLa, or NIH3T3) are cultured in methionine-free DMEM supplemented with 10% dialyzed FBS. L-Azidohomoalanine (50-100 microM) is added to the medium for 2-24 hours to label newly synthesized proteins. Cells are then fixed, permeabilized, and reacted with DBCO-fluorophore or DBCO-biotin for click chemistry labeling. Labeled proteins are visualized by fluorescence microscopy or detected by Western blot with streptavidin-HRP. For proteomic analysis, labeled proteins are enriched with biotin-avidin and analyzed by LC-MS/MS.
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| Animal Protocol |
In vivo animal experiments for L-Azidohomoalanine involve administration to mice for protein labeling. C57BL/6 mice receive intraperitoneal injection of L-Azidohomoalanine (10-50 mg/kg) dissolved in PBS. After 1-24 hours, mice are euthanized, and tissues (brain, liver, kidney, muscle) are collected, homogenized, and subjected to click chemistry with DBCO-biotin or DBCO-fluorophore. Labeled proteins are analyzed by Western blot, fluorescence imaging, or mass spectrometry. This approach enables measurement of protein synthesis rates and identification of newly synthesized proteins in vivo. All procedures comply with institutional animal care guidelines.
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| References |
| Molecular Formula |
C4H8N4O2
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|---|---|
| Molecular Weight |
144.13192
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| Exact Mass |
144.065
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| CAS # |
120042-14-0
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| Related CAS # |
120042-14-0;942518-29-8 (HCl);
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| PubChem CID |
147480
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| Appearance |
Typically exists as solid at room temperature
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| Melting Point |
220-223ºC
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| LogP |
0.251
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
10
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| Complexity |
163
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| Defined Atom Stereocenter Count |
1
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| SMILES |
[N-]=[N+]=NCC[C@@H](C(=O)O)N
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| InChi Key |
NNWQLZWAZSJGLY-VKHMYHEASA-N
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| InChi Code |
InChI=1S/C4H8N4O2/c5-3(4(9)10)1-2-7-8-6/h3H,1-2,5H2,(H,9,10)/t3-/m0/s1
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| Chemical Name |
(2S)-2-amino-4-azidobutanoic acid
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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 |
| 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 | 6.9382 mL | 34.6909 mL | 69.3818 mL | |
| 5 mM | 1.3876 mL | 6.9382 mL | 13.8764 mL | |
| 10 mM | 0.6938 mL | 3.4691 mL | 6.9382 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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