| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
3-Azido-L-alanine functions as a chemical reporter incorporated into microcystins via precursor-directed biosynthesis for bioorthogonal labeling, without direct interaction with biological receptors or enzymes [1]
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|---|---|
| ln Vitro |
- Biosynthetic Incorporation into Microcystins:
1. Cyanobacterial Cultivation: 3-Azido-L-alanine (0.1–1 mM) was supplemented into BG-11 medium for Microcystis aeruginosa cultures. Incorporation efficiency was assessed via LC-MS, showing 3-azidoalanine substitution at the Adda moiety position (mass shift +28 Da) in 20–40% of total microcystins after 7 days [1] 2. Click Chemistry Labeling: Azido-functionalized microcystins were reacted with DBCO-fluorophores (10 μM, 37°C, 1 h) in PBS buffer, achieving >90% labeling efficiency as confirmed by fluorescence spectroscopy (λex 488 nm, λem 520 nm) [1] - Cytotoxicity Retention: |
| ln Vivo |
- Cellular Uptake Dynamics:
1. Time-Lapse Microscopy: Fluorescent microcystins (10 nM) were internalized by HepG2 cells expressing OATP1B1/1B3 transporters within 5 minutes, accumulating in perinuclear vesicles. Uptake was blocked by 10 μM rifampicin (OATP inhibitor) [1] |
| Enzyme Assay |
- Protein Phosphatase Inhibition Assay [1]:
1. Reaction Setup: Recombinant PP1 catalytic subunit (20 nM) was incubated with 32P-labeled phosphorylase a (1 μM) in assay buffer (50 mM Tris-HCl pH 7.5, 10 mM MgCl₂). 3-Azido-L-alanine-labeled microcystin (0.1–100 nM) was added, and dephosphorylation was stopped with SDS-PAGE loading buffer after 30 minutes at 37°C. 2. Quantification: Phosphorylase a activity was measured by autoradiography, showing IC₅₀ values identical to native microcystin-LR (8 ± 1 nM), confirming azido substitution did not impair enzyme binding [1] |
| Cell Assay |
- Microcystin Incorporation and Labeling Protocol [1]:
1. Cyanobacterial Culture: Microcystis aeruginosa cells were grown in BG-11 medium supplemented with 3-Azido-L-alanine (0.5 mM) for 7 days. Cells were harvested by centrifugation (5,000g, 10 min), and microcystins were extracted with 80% methanol. 2. Click Reaction: Extracts were reacted with DBCO-PEG4-TAMRA (10 μM) in PBS (pH 7.4) for 1 h at room temperature. Labeled microcystins were purified by C18 SPE cartridges and analyzed by LC-MS/MS - Confocal Microscopy: 1. HepG2 Cell Staining: Cells treated with labeled microcystins (10 nM) were fixed with 4% PFA, permeabilized with 0.1% Triton X-100, and counterstained with DAPI. Fluorescence colocalization with LysoTracker Red (lysosomal marker) confirmed vesicular trafficking [1] |
| References | |
| Additional Infomation |
Biosynthetic Engineering:
1. Precursor Specificity: The incorporation capacity of Microcystis strains varies, with Microcystis aeruginosa PCC 7806 showing higher tolerance (up to 1 mM) than Microcystis. (Maximum concentration 0.2 mM) [1] 2. Metabolic competition: Incorporation efficiency is improved by simultaneously supplementing L-alanine (0.5 mM), thereby reducing competition with the natural amino acid library [1] - Application scope: 1. Imaging probe development: Clickable microcystin can be used for live cell tracking of toxin transport, revealing OATP-dependent uptake pathways [1] 2. Structure-activity studies: Azide substitution allows for site-specific coupling of affinity tags for pull-down experiments, thereby identifying new protein-protein interactors [1] - Limitations: 1. Low natural abundance: After supplementation with azide alanine, the endogenous microcystin yield in Microcystis decreased by 30-50%, which may be due to metabolic stress [1] |
| Molecular Formula |
C3H6N4O2
|
|---|---|
| Molecular Weight |
130.11
|
| Exact Mass |
130.049
|
| CAS # |
105661-40-3
|
| Related CAS # |
3-Azido-L-alanine hydrochloride;1620171-64-3
|
| PubChem CID |
9964096
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| Appearance |
Typically exists as solid at room temperature
|
| LogP |
-2.5
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
9
|
| Complexity |
150
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
N(C[C@@H](C(O)=O)N)=[N+]=[N-]
|
| InChi Key |
CIFCKCQAKQRJFC-REOHCLBHSA-N
|
| InChi Code |
InChI=1S/C3H6N4O2/c4-2(3(8)9)1-6-7-5/h2H,1,4H2,(H,8,9)/t2-/m0/s1
|
| Chemical Name |
(2S)-2-amino-3-azidopropanoic acid
|
| Synonyms |
3-Azido-L-alanine; 105661-40-3; L-Alanine, 3-azido-; L-Azidoalanine; ALANINE, 3-AZIDO-, L-; UNII-8Y835MJ6LI; 8Y835MJ6LI; CCRIS 1790;
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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
|
|---|---|
| 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 | 7.6858 mL | 38.4290 mL | 76.8580 mL | |
| 5 mM | 1.5372 mL | 7.6858 mL | 15.3716 mL | |
| 10 mM | 0.7686 mL | 3.8429 mL | 7.6858 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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