| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
L-ANAP targets specific sites within proteins when incorporated during translation. It is not a drug targeting a disease-related receptor or enzyme, but rather a tool for imaging and studying protein conformations by sensing changes in the local polarity of the environment.
|
|---|---|
| ln Vitro |
For particular fluorescent labeling, L-ANAP, a fluorescent noncanonical amino acid, can be employed[1]. L-ANAP is a fluorescent unnatural amino acid (Uaa) that is polarity-sensitive and genetically encodable. Molecular and cellular biology are particularly interested in fluorescent Uaas, such as L-dansylalanine, L-(7-hydroxycoumrin-4-yl)ethylglycine, and L-ANAP, which may enhance and supplement the commonly employed fluorescent proteins (FPs)[2].
L-ANAP is a polarity-sensitive fluorescent unnatural amino acid. The fluorescence properties of L-ANAP change in response to the hydrophobicity or polarity of its local environment, allowing researchers to monitor protein conformational changes, protein-protein interactions, and local structural dynamics in live cells. |
| ln Vivo |
L-ANAP is not administered in vivo as a drug or therapeutic agent. When genetically incorporated into proteins expressed in cells, it enables the study of protein folding, dynamics, and interactions in living systems. It has been used to study conformational changes of receptors and ion channels.
|
| Enzyme Assay |
Non-cell binding assays for L-ANAP are not based on receptor-ligand interactions. Instead, the compound is used to label purified proteins of interest. Recombinant proteins containing L-ANAP are expressed in E. coli or mammalian cells using genetic code expansion technology. The purified protein is then analyzed by fluorescence spectroscopy to characterize its emission properties. Typically, fluorescence excitation is performed at 360-400 nm, and emission spectra are recorded between 400-600 nm. The sensitivity to polarity is assessed by measuring changes in fluorescence intensity or shift in emission maximum (λmax) in response to changes in solvent conditions (e.g., varying concentration of denaturants like guanidine hydrochloride or urea, or changes in pH). Time-resolved fluorescence measurements can be performed to measure fluorescence lifetimes, which are also sensitive to the local environment. The polarity sensitivity allows L-ANAP to function as a local probe of protein structure, reporting on the hydrophobicity of the amino acid's surrounding environment with high spatial resolution.
|
| Cell Assay |
Cellular assays for L-ANAP are performed using mammalian cell lines such as HEK293, HeLa, or CHO cells, or bacterial cells such as E. coli. For mammalian cells, an orthogonal tRNA/aminoacyl-tRNA synthetase pair (e.g., from Methanococcus jannaschii or Methanosarcina mazei) is co-transfected with a plasmid encoding the gene of interest containing an amber stop codon (TAG) at a specific position. Cells are cultured in medium supplemented with 0.1-1 mM L-ANAP hydrochloride for 24-48 hours. After expression, cells are washed with PBS, and fluorescence is visualized by confocal or widefield fluorescence microscopy using appropriate filter sets (excitation ~400 nm, emission ~450-550 nm). For live-cell imaging, cells are maintained in phenol red-free medium at 37degC with 5% CO2. For flow cytometry analysis, cells are trypsinized, resuspended in PBS, and analyzed on a flow cytometer equipped with a violet laser (405 nm excitation) and appropriate emission filters. For protein interaction studies, fluorescence resonance energy transfer (FRET) can be measured if L-ANAP is incorporated adjacent to another fluorophore or if used in combination with fluorescent proteins. For conformational studies, changes in L-ANAP fluorescence in response to addition of ligands, changes in temperature, or other stimuli are measured using a fluorescence plate reader.
|
| Animal Protocol |
Animal studies with L-ANAP are primarily conducted in invertebrate model organisms such as C. elegans or zebrafish, where genetic code expansion techniques have been established for incorporating unnatural amino acids into proteins expressed in specific tissues. For zebrafish, one-cell stage embryos are microinjected with mRNA encoding the engineered tRNA and aminoacyl-tRNA synthetase, along with mRNA or plasmid DNA encoding the target protein with an amber codon, in the presence of L-ANAP hydrochloride (0.1-1 mM) in the injection buffer. Embryos are incubated in fish water containing L-ANAP (0.1-0.5 mM) for 24-72 hours. Fluorescence imaging is performed on a confocal or widefield fluorescence microscope equipped with water-immersion objectives. For C. elegans, animals are grown on nematode growth medium (NGM) agar plates seeded with E. coli expressing the target protein or supplemented with L-ANAP-containing media, and fluorescence is observed in live animals using fluorescence microscopy. Mammalian studies are limited due to challenges with systemic delivery of unnatural amino acids and the requirements for organ-specific expression of the orthogonal translation machinery, though transgenic mouse models expressing the orthogonal tRNA/synthetase pair have been developed for tissue-specific unnatural amino acid incorporation.
|
| ADME/Pharmacokinetics |
L-ANAP hydrochloride is a small fluorescent unnatural amino acid (molecular weight 308.76 for the hydrochloride salt) that is supplied as a research chemical. Pharmacokinetic properties in animals are not well characterized because L-ANAP is not administered as a therapeutic agent. When added to cell culture medium (typically 0.1-1 mM), L-ANAP is taken up by cells through amino acid transporters such as LAT1 (SLC7A5) and system L transporters. Once inside cells, it is incorporated into proteins during translation by the orthogonal tRNA/synthetase pair. Unincorporated L-ANAP is presumed to be metabolized or excreted via normal amino acid catabolic pathways and renal clearance. The compound is stable in cell culture medium at 37degC for 24-48 hours. Detailed parameters such as half-life, volume of distribution, and plasma protein binding have not been established because L-ANAP is not intended for pharmaceutical development.
|
| Toxicity/Toxicokinetics |
L-ANAP hydrochloride is a fluorescent unnatural amino acid used for research purposes only and not as a therapeutic agent; therefore, no formal toxicity studies have been conducted. Based on its chemical structure, L-ANAP hydrochloride is expected to have low acute toxicity when handled appropriately with standard laboratory precautions. The compound should be treated as a potential irritant; safety data sheets (SDS) recommend avoiding inhalation, ingestion, and skin contact. When used in cell culture at typical concentrations (0.1-1 mM), L-ANAP does not exhibit significant cytotoxicity or adverse effects on cell viability, as determined by standard assays (MTT, CellTiter-Glo) in HEK293, HeLa, and CHO cells. Long-term toxicity studies have not been performed. In invertebrate models such as C. elegans and zebrafish, L-ANAP has been used at concentrations up to 0.5-1 mM without reports of gross toxicity or developmental abnormalities. The hydrochloride salt may cause mild irritation upon direct contact with mucous membranes. No mutagenicity or genotoxicity data are available for L-ANAP hydrochloride.
|
| References |
|
| Additional Infomation |
L-ANAP (Amino-Naphthalene-Alanine-Phenylalanine, or more specifically, 3-(6-acetylnaphthalen-2-ylamino)-2-aminopropanoic acid) is a fluorescent unnatural amino acid that is genetically encodable and polarity-sensitive, meaning its fluorescence properties change based on the hydrophobicity of its local environment. The key advantage of L-ANAP over traditional fluorescent proteins is its small size and ability to be placed at specific, defined positions within a protein of interest with minimal perturbation to protein structure. This allows researchers to study protein conformational changes at near-atomic resolution, monitor protein folding pathways in real-time, track protein-protein interactions, and map binding sites for ligands or other proteins. L-ANAP has been used to study conformational dynamics of G protein-coupled receptors (GPCRs), ion channels (including voltage-gated potassium channels), and enzymes. The excitation maximum is approximately 360-400 nm, and the emission maximum ranges from 450-550 nm depending on the polarity of the environment. L-ANAP is part of a growing toolkit of fluorescent unnatural amino acids that supplement commonly used fluorescent proteins (FPs), offering the ability to study protein structure and dynamics with higher spatial resolution and less steric hindrance. The compound is strictly for research use and is not approved for clinical or diagnostic applications.
|
| Molecular Formula |
C15H17CLN2O3
|
|---|---|
| Molecular Weight |
308.76
|
| Related CAS # |
L-ANAP;1313516-26-5
|
| Appearance |
White to yellow solid powder
|
| 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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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) |
DMSO :~33.33 mg/mL (~107.95 mM)
H2O :~1 mg/mL (~3.24 mM) |
|---|---|
| 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 | 3.2388 mL | 16.1938 mL | 32.3876 mL | |
| 5 mM | 0.6478 mL | 3.2388 mL | 6.4775 mL | |
| 10 mM | 0.3239 mL | 1.6194 mL | 3.2388 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
