| Size | Price | Stock | Qty |
|---|---|---|---|
| 100mg |
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| 500mg |
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| Other Sizes |
|
| Targets |
Click chemistry reagent; unnatural amino acid
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|---|---|
| ln Vitro |
Modified soluble superfolded green fluorescent protein (sfGFP) containing 4-azido-L-phenylalanine or p-propargyloxy-L-phenylalanine (pPaF) will accumulate 0.9-1.7 mg/mL in CFPS solutions when using a cell-free protein synthesis (CFPS) approach [1].
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| Enzyme Assay |
Twenty-five-microlitre PANOx SP reaction solutions contained, unless otherwise noted: 10 mM ammonium glutamate, 20 mM magnesium glutamate, 175 mM potassium glutamate, 1.2 mM ATP, 0.86 mM each of CMP, GMP and UMP, 10 mM dibasic potassium phosphate, 34 μg/ml folinic acid, 171 μg/ml E. coli tRNAs, 33 mM phosphoenolpyruvate (PEP), 1.5 mM spermidine, 1 mM putrescine, 0.33 mM nicotinamide adenine dinucleotide (NAD), 0.27 mM coenzyme A, 2.7 mM sodium oxalate, 2 mM each of the 20 amino acids, 5 μM l-[14C(U)]-leucine, 2 mM p-azido-l-phenylalanine (pAzF) or 4 mM p-propargyloxy-l-phenylalanine (pPaF), 0.3 mg/ml pAzFRS or 0.5 mg/ml pPaFRS (prepared as described above), 100 μg/ml T7 RNA polymerase, 6 nM sfGFP plasmid, 0.2 mg/ml linearized o-tRNA or o-tRNAopt plasmid template and either 0.24 volume of standard KC6 extract or 0.28 volume of orthogonal extract. The glutamate salts were diluted from a 10-fold concentrated (10×) solution. Similarly, a 10× NTMP Master Mix solution contained ATP, the three NMPs, folinic acid and the E. coli tRNAs, and was adjusted to pH 7.3 with dibasic potassium phosphate. The natural amino acids were diluted from a stock solution, which contained a 50 mM concentration of each amino acid. This stock solution was prepared by adding the amino acids in the following order (given in their one-letter code): R, V, W, F, I, L, C, M, A, N, D, E, G, Q, H, K, P, S, T, Y. During the preparation, it was ensured that each amino acid was dissolved before addition of the next, except tyrosine, which is added last and remains suspended in the solution. The reactions were incubated at 30°C for 16 h, unless stated otherwise. CFPS reactions containing pAzF were prepared in a dark room, and the tubes containing the reaction solutions were wrapped in aluminum foil to prevent photodissociation of the aromatic azide[1].
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| Cell Assay |
The orthogonal S30 cell extract was prepared from KC6 cells harbouring the pDule-tRNA plasmid, as described. The lysate was not diluted after homogenization. The specific growth rate of this KC6/o-tRNA culture was 0.57 h−1 during the batch fermentation. The standard KC6 extract was prepared as described. After homogenization, 0.2 ml S30 buffer (14 mM magnesium acetate, 60 mM potassium acetate, 10 mM Tris acetate, pH 8.2) was added per ml of lysate, and the diluted lysate was dialysed against 100 volumes of S30 buffer at 4°C four times for 30 min each. Finally, the S30 cell extract was centrifuged at 10 000g and 4°C for 20 min, and the supernatant was aliquoted, flash-frozen and stored at −80°C. The specific growth rate of the KC6 culture was 0.96 h−1 during the fed-batch fermentation[1].
|
| References |
[1]. Albayrak C, et al. Cell-free co-production of an orthogonal transfer RNA activates efficient site-specific non-natural amino acid incorporation. Nucleic Acids Res. 2013 Jun;41(11):5949-63.
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| Molecular Formula |
C9H11CLN4O2
|
|---|---|
| Molecular Weight |
242.6622402668
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| Exact Mass |
242.057
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| Elemental Analysis |
C, 44.55; H, 4.57; Cl, 14.61; N, 23.09; O, 13.19
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| CAS # |
34670-43-4
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| Related CAS # |
4-Azido-L-phenylalanine;33173-53-4
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| PubChem CID |
86585254
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| Appearance |
Typically exists as off-white to light yellow solids at room temperature
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
16
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| Complexity |
267
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| Defined Atom Stereocenter Count |
1
|
| SMILES |
C(O)(=O)[C@H](CC1=CC=C(N=[N+]=[N-])C=C1)N.[H]Cl
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| InChi Key |
VXCXRGSRHCHMBK-QRPNPIFTSA-N
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| InChi Code |
InChI=1S/C9H10N4O2.ClH/c10-8(9(14)15)5-6-1-3-7(4-2-6)12-13-11;/h1-4,8H,5,10H2,(H,14,15);1H/t8-;/m0./s1
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| Chemical Name |
(2S)-2-amino-3-(4-azidophenyl)propanoic acid;hydrochloride
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| Synonyms |
34670-43-4; 4-Azido-L-phenylalanine hydrochloride; 4-Azido-L-phenylalanine (hydrochloride); (2S)-2-amino-3-(4-azidophenyl)propanoic acid;hydrochloride; (2S)-2-AMINO-3-(4-AZIDOPHENYL)PROPANOIC ACID HYDROCHLORIDE; 4-Azido-L-phenylalanine HCl; 3-(p-Azidophenyl)-L-alanine Hydrochloride; 4-Azidophenylalanine Hydrochloride; p-Azido-L-phenylalanine Hydrochloride; p-Azidophenylalanine Hydrochloride; p-Azido-L-phenylalanine hydrochloride;
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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) |
DMSO : ≥ 103.3 mg/mL (~425.70 mM)
H2O : ~10 mg/mL (~41.21 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (10.30 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 25.0 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: ≥ 2.5 mg/mL (10.30 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 25.0 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: ≥ 2.5 mg/mL (10.30 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: 16.67 mg/mL (68.70 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication (<60°C). |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.1210 mL | 20.6050 mL | 41.2099 mL | |
| 5 mM | 0.8242 mL | 4.1210 mL | 8.2420 mL | |
| 10 mM | 0.4121 mL | 2.0605 mL | 4.1210 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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