| Size | Price | Stock | Qty |
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| 250mg |
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| 500mg |
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| 1g |
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| 2g |
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| 5g |
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| Other Sizes |
| Targets |
L-Leucine-p-nitroanilide serves as a substrate for leucine aminopeptidase (LAP) and other aminopeptidases. These enzymes catalyze the hydrolysis of the peptide bond between leucine and p-nitroaniline, releasing free leucine and the chromogenic p-nitroaniline. The compound does not have a therapeutic target per se but is used as a tool to measure the activity of these enzymes. Leucine aminopeptidase is involved in protein turnover and is found in various tissues; altered LAP activity is associated with certain liver diseases and cancers.
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|---|---|
| ln Vitro |
In vitro, L-Leucine-p-nitroanilide is used as a substrate to measure leucine aminopeptidase activity. The enzyme cleaves the compound, releasing p-nitroaniline, which can be quantified by measuring absorbance at 405 nm. The rate of p-nitroaniline production is directly proportional to enzyme activity. This assay is used to determine the kinetic parameters of aminopeptidases (Km, Vmax) and to screen for enzyme inhibitors. The compound is also used in clinical chemistry for the measurement of LAP activity in serum or other biological fluids.
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| ln Vivo |
In vivo, L-Leucine-p-nitroanilide is not used as a therapeutic agent. It is primarily a research and diagnostic tool. The compound may be used in animal studies to assess aminopeptidase activity in tissues or to evaluate the efficacy of aminopeptidase inhibitors. However, detailed in vivo activity data are not extensively reported in the available literature. Its primary application remains in in vitro enzyme assays.
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| Enzyme Assay |
Non-cell-based enzyme assays for L-Leucine-p-nitroanilide involve incubating the substrate with a sample containing leucine aminopeptidase (e.g., purified enzyme, tissue homogenate, or serum) in a suitable buffer at physiological pH. The reaction is incubated at 37°C for a specified time, and the reaction is stopped (e.g., by adding acetic acid). The absorbance of the released p-nitroaniline is measured at 405 nm using a spectrophotometer. Enzyme activity is calculated from a standard curve of p-nitroaniline. Kinetic parameters can be determined by varying substrate concentrations.
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| Cell Assay |
Cellular assays for L-Leucine-p-nitroanilide are not typically performed, as the compound is used as a substrate in cell-free enzyme assays. However, it may be used to measure aminopeptidase activity in cell lysates or culture supernatants. Cells are lysed, and the lysate is incubated with the substrate as described above. Alternatively, cell-based assays may use the compound to assess the activity of cell surface aminopeptidases, but detailed protocols are not extensively reported.
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| Animal Protocol |
In vivo animal models for L-Leucine-p-nitroanilide are not typical, as the compound is a research and diagnostic tool rather than a therapeutic agent. It may be used in pharmacokinetic studies to assess the stability of the compound in biological fluids or to measure aminopeptidase activity in tissue homogenates from animal models of disease. Detailed protocols are not extensively reported in the available literature.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of L-Leucine-p-nitroanilide include CAS number 4178-93-2. The compound is a small molecule dipeptide analog. It is soluble in appropriate buffers for enzymatic assays. Detailed PK parameters such as half-life, bioavailability, and volume of distribution are not extensively reported, as the compound is not used as a therapeutic agent. It is a research and diagnostic reagent.
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| Toxicity/Toxicokinetics |
L-Leucine-p-nitroanilide is a chromogenic substrate with generally low toxicity in the context of its use as a research reagent. Detailed toxicological data are not extensively reported, as the compound is not intended for therapeutic use. Standard laboratory safety precautions should be followed when handling the compound. It is for research use only and not for human consumption.
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| Additional Infomation |
L-Leucine-p-nitroanilide is also known as Leu-pNA and L-Leu-pNA. It has CAS number 4178-93-2. It is a chromogenic substrate for leucine aminopeptidase and other aminopeptidases. Upon enzymatic cleavage, it releases p-nitroaniline, which can be quantified spectrophotometrically at 405 nm. It is used for enzyme activity assays, enzyme kinetics studies, and inhibitor screening in biochemical and clinical research. It is for research use only.
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| Molecular Formula |
C12H17N3O3
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|---|---|
| Molecular Weight |
251.282
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| Exact Mass |
251.126
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| CAS # |
4178-93-2
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| PubChem CID |
2733285
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.2±0.1 g/cm3
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| Boiling Point |
454.5±30.0 °C at 760 mmHg
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| Melting Point |
88-90ºC
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| Flash Point |
228.7±24.6 °C
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| Vapour Pressure |
0.0±1.1 mmHg at 25°C
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| Index of Refraction |
1.589
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| LogP |
1.83
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
18
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| Complexity |
293
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CC(C)C[C@@H](C(=O)NC1=CC=C(C=C1)[N+](=O)[O-])N
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| InChi Key |
AXZJHDNQDSVIDR-NSHDSACASA-N
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| InChi Code |
InChI=1S/C12H17N3O3/c1-8(2)7-11(13)12(16)14-9-3-5-10(6-4-9)15(17)18/h3-6,8,11H,7,13H2,1-2H3,(H,14,16)/t11-/m0/s1
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| Chemical Name |
(2S)-2-amino-4-methyl-N-(4-nitrophenyl)pentanamide
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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 | 3.9796 mL | 19.8981 mL | 39.7962 mL | |
| 5 mM | 0.7959 mL | 3.9796 mL | 7.9592 mL | |
| 10 mM | 0.3980 mL | 1.9898 mL | 3.9796 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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