| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
26S proteasome (caspase-like site / post-glutamyl peptide hydrolase). Ac-Nle-Pro-Nle-Asp-AMC is a specific peptide substrate for the caspase-like activity of the proteasome.
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| ln Vitro |
Ac-Nle-Pro-Nle-Asp-AMC is cleaved by the caspase-like site of the 26S proteasome with specific activities of 113 nmol/min/mg for rabbit muscle 26S proteasome and 6.6 nmol/min/mg for yeast 20S proteasome. The AMC moiety enables sensitive fluorescence detection of proteolytic cleavage.
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| ln Vivo |
Ac-Nle-Pro-Nle-Asp-AMC is not a therapeutic agent but a research tool for measuring proteasome activity ex vivo. It has been used in animal tissue lysates to assess proteasome function in disease models, including studies of cancer, neurodegeneration, and muscle wasting, where proteasome activity is dysregulated.
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| Enzyme Assay |
For non-cellular assays, purified 20S or 26S proteasome is incubated in assay buffer containing 50 mM Tris-HCl (pH 7.5), 1 mM DTT, and 5 mM MgCl2. Ac-Nle-Pro-Nle-Asp-AMC substrate is added at 50-100 microM final concentration. The reaction is incubated at 37degC for 30-60 min, and AMC fluorescence is measured at excitation 380 nm and emission 460 nm using a plate reader.
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| Cell Assay |
Ac-Nle-Pro-Nle-Asp-AMC is typically not used directly in cell-based assays as a treatment; rather, cells are treated with drugs to modulate proteasome activity, then lysed. The lysate is incubated with the substrate to measure caspase-like proteasome activity. Results are normalized to protein concentration and/or expressed as fold-change relative to control.
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| Animal Protocol |
For ex vivo activity measurement, animals are treated with test compounds, then tissues (liver, muscle, kidney, tumor) are harvested and homogenized in lysis buffer. Lysates are clarified by centrifugation, and protein concentration is quantified. Lysates are then incubated with Ac-Nle-Pro-Nle-Asp-AMC (100 microM) for 30-60 min at 37degC, and fluorescence is measured to determine proteasome caspase-like activity.
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| ADME/Pharmacokinetics |
Ac-Nle-Pro-Nle-Asp-AMC itself has no significant pharmacokinetics as a molecule of interest; its stability in assay buffers (pH 7-8) is sufficient for standard proteasome activity measurements. It should be stored at -20degC, protected from light, and reconstituted in DMSO for a stable stock solution.
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| Toxicity/Toxicokinetics |
Toxicity of Ac-Nle-Pro-Nle-Asp-AMC is not relevant as it is not used in vivo therapeutically. Standard laboratory safety precautions for handling fluorescent peptides should be followed, including avoiding inhalation and skin contact. It is intended for research use only.
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| References |
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| Additional Infomation |
Ac-Nle-Pro-Nle-Asp-AMC (CAS: 355140-49-7) has molecular formula C33H45N5O9 and molecular weight 655.74. The sequence is Acetyl-Norleucine-Proline-Norleucine-Aspartic acid-AMC. It is a valuable tool for dissecting the contribution of the caspase-like site to overall proteasome function, particularly in drug development for proteasome inhibitors such as bortezomib.
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| Molecular Formula |
C33H45N5O9
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|---|---|
| Molecular Weight |
655.74
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| Exact Mass |
655.321
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| CAS # |
355140-49-7
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| PubChem CID |
131698166
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| Appearance |
White to off-white solid powder
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
902.2±75.0 °C at 760 mmHg
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| Flash Point |
499.4±37.1 °C
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| Vapour Pressure |
0.0±0.3 mmHg at 25°C
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| Index of Refraction |
1.617
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| LogP |
6.08
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
16
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| Heavy Atom Count |
47
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| Complexity |
1220
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| Defined Atom Stereocenter Count |
4
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| SMILES |
CCCC[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)NC1=CC2=C(C=C1)C(=CC(=O)O2)C)NC(=O)[C@@H]3CCCN3C(=O)[C@H](CCCC)NC(=O)C
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| InChi Key |
WPTAROWBMZVECB-CQJMVLFOSA-N
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| InChi Code |
InChI=1S/C33H45N5O9/c1-5-7-10-23(36-32(45)26-12-9-15-38(26)33(46)24(11-8-6-2)34-20(4)39)30(43)37-25(18-28(40)41)31(44)35-21-13-14-22-19(3)16-29(42)47-27(22)17-21/h13-14,16-17,23-26H,5-12,15,18H2,1-4H3,(H,34,39)(H,35,44)(H,36,45)(H,37,43)(H,40,41)/t23-,24-,25-,26-/m0/s1
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| Chemical Name |
(3S)-3-[[(2S)-2-[[(2S)-1-[(2S)-2-acetamidohexanoyl]pyrrolidine-2-carbonyl]amino]hexanoyl]amino]-4-[(4-methyl-2-oxochromen-7-yl)amino]-4-oxobutanoic acid
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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: ≥ 100 mg/mL (152.50 mM)
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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 | 1.5250 mL | 7.6250 mL | 15.2499 mL | |
| 5 mM | 0.3050 mL | 1.5250 mL | 3.0500 mL | |
| 10 mM | 0.1525 mL | 0.7625 mL | 1.5250 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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