| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| Other Sizes |
| Targets |
Z-Nle-Lys-Arg-AMC acetate targets cathepsin B, a cysteine protease that is involved in intracellular protein degradation, antigen processing, and extracellular matrix remodeling. Cathepsin B is overexpressed in various cancers and is associated with tumor invasion, metastasis, and angiogenesis. It is also implicated in inflammatory diseases and neurodegenerative disorders such as Alzheimer's disease. The substrate's sequence (Nle-Lys-Arg) is specifically recognized and cleaved by cathepsin B.
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| ln Vitro |
In vitro, Z-Nle-Lys-Arg-AMC acetate is used as a fluorogenic substrate to measure cathepsin B enzymatic activity in biochemical assays. The substrate is cleaved by active cathepsin B, releasing the fluorescent AMC moiety, which can be detected at excitation/emission wavelengths of approximately 355/460 nm. The increase in fluorescence over time is directly proportional to enzyme activity. The substrate can be used to determine enzyme kinetics (Km, Vmax), inhibitor potency (IC50), and to screen for cathepsin B inhibitors in drug discovery programs.
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| ln Vivo |
In vivo activity is not directly measured for Z-Nle-Lys-Arg-AMC acetate, as it is a substrate used in biochemical assays rather than a therapeutic agent. However, the substrate can be used to assess cathepsin B activity in tissue homogenates, plasma, or other biological samples collected from animal models of disease. Elevated cathepsin B activity detected using this substrate can serve as a biomarker of disease progression or treatment response in preclinical studies.
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| Enzyme Assay |
Non-cellular enzyme assays using Z-Nle-Lys-Arg-AMC acetate are performed in buffer systems optimized for cathepsin B activity. The substrate is incubated with purified cathepsin B enzyme or tissue lysates at the appropriate pH (typically pH 5.5-6.0 for cathepsin B). The reaction is monitored continuously using a fluorescence plate reader, and the initial rate of fluorescence increase is calculated. Inhibitors are added to the reaction to determine their potency. Controls including enzyme blanks and substrate blanks are included to account for background fluorescence.
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| Cell Assay |
In vitro cellular experiments using Z-Nle-Lys-Arg-AMC acetate are limited due to the substrate's inability to cross cell membranes. However, cell lysates can be prepared from treated cells, and cathepsin B activity in the lysates can be measured using the substrate. This approach allows researchers to assess the effects of test compounds on cathepsin B activity in cellular contexts. Cells are treated with compounds of interest, lysed, and the lysates are incubated with the substrate to measure enzymatic activity.
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| Animal Protocol |
In vivo animal experiments using Z-Nle-Lys-Arg-AMC acetate involve collecting tissue samples or biological fluids from animals and measuring cathepsin B activity ex vivo. The substrate is incubated with the samples, and fluorescence is measured to quantify enzyme activity. This approach can be used to monitor disease progression or evaluate the pharmacodynamic effects of cathepsin B inhibitors in preclinical models of cancer, inflammation, or neurodegeneration.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties are not relevant for Z-Nle-Lys-Arg-AMC acetate as it is a substrate used in biochemical assays and not a therapeutic compound. The molecular weight is approximately 782.9 g/mol. The compound is supplied as an acetate salt. It should be stored according to the manufacturer's recommendations, typically at -20°C protected from light. The substrate should be dissolved in an appropriate buffer or solvent for use in enzymatic assays.
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| Toxicity/Toxicokinetics |
Toxicity data for Z-Nle-Lys-Arg-AMC acetate are not typically characterized as the compound is a research reagent used in enzymatic assays, not a therapeutic agent. Standard laboratory safety precautions should be followed when handling the compound. The AMC fluorophore is generally considered to have low toxicity in the context of in vitro assays. Researchers should consult the safety data sheet for specific handling and disposal guidelines.
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| References | |
| Additional Infomation |
Z-Nle-Lys-Arg-AMC acetate is a fluorogenic peptide substrate for cathepsin B. It is also known as Z-Nle-Lys-Arg-AMC (acetate). The substrate is used to monitor cathepsin B activity over a wide pH range. It is widely applied in cancer, inflammation, and neurodegeneration research. The substrate is for research use only and not for human therapeutic applications.
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| Molecular Formula |
C36H50N8O7.XC2H4O2
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| Molecular Weight |
706.83 (free base)
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| Appearance |
White to off-white solid powder
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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 (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)
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| Solubility (In Vitro) |
H2O :≥ 100 mg/mL
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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.) |
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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