| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| 100mg | |||
| Other Sizes |
| Targets |
Z-Ala-Ala-Asn-AMC targets legumain, a cysteine protease also known as asparaginyl endopeptidase (AEP). Legumain cleaves the peptide bond after asparagine residues. The compound serves as a substrate for legumain, and its cleavage releases the fluorescent AMC group. It does not have a therapeutic target but is a research tool for measuring legumain activity in vitro and in biological samples.
|
|---|---|
| ln Vitro |
The P3-P2-P1 sequence Ala-Ala-Asn served as the basis for the first legumain fluorescent substrate, Cbz-Ala-Asn-AMC. S effectively cleaves the synthetic substrate Cbz-Ala-Ala-Asn-AMC. Human and mansoni legumin, respectively, have Kms of 80 and 90 μM[2][3].
In vitro, Z-Ala-Ala-Asn-AMC is used as a fluorogenic substrate to measure legumain activity. The compound is incubated with a sample containing legumain, and the release of AMC is monitored by fluorescence (excitation at 350-380 nm, emission at 450-460 nm). The rate of fluorescence increase is proportional to enzyme activity. It is used to study legumain activity in various biological contexts. |
| ln Vivo |
Z-Ala-Ala-Asn-AMC is not a pharmacologically active compound and therefore does not have defined in vivo activity as a therapeutic agent. It is a research tool used to measure legumain activity in biological samples. When administered to animals, it would likely be cleaved by legumain to release the AMC fluorophore. Its primary value is as a diagnostic and research reagent for studying legumain activity in health and disease.
|
| Enzyme Assay |
In vitro enzyme assays for Z-Ala-Ala-Asn-AMC are designed to measure legumain activity. A standard protocol involves incubating the substrate with a sample containing legumain (e.g., tissue homogenate, cell lysate, or purified enzyme) in a suitable buffer at 37°C. The release of AMC is monitored continuously or at fixed time points using a fluorometer with excitation at 350-380 nm and emission at 450-460 nm. The initial velocity is calculated, and the enzyme activity is determined.
|
| Cell Assay |
In vitro cellular assays using Z-Ala-Ala-Asn-AMC are conducted to measure legumain activity in cell cultures. Cells are lysed, and the lysate is incubated with the substrate. The release of AMC is measured fluorometrically. These assays are used to study the regulation of legumain expression and activity in various cell types and to screen for compounds that modulate legumain activity.
|
| Animal Protocol |
In vivo animal studies with Z-Ala-Ala-Asn-AMC are not typically conducted for therapeutic purposes. However, it may be used in pharmacokinetic or pharmacodynamic studies to measure legumain activity in vivo. A typical protocol involves administering the substrate to rodents, followed by blood sampling to measure the rate of AMC release. These studies help to assess the in vivo activity of legumain and the efficacy of legumain inhibitors.
|
| ADME/Pharmacokinetics |
As a small, hydrophilic peptide substrate, Z-Ala-Ala-Asn-AMC is expected to be rapidly cleared from the circulation. Its pharmacokinetic properties are characteristic of small peptides, with rapid clearance and short half-life. The compound is used primarily as a research reagent for in vitro assays, and detailed pharmacokinetic data are not typically reported.
|
| Toxicity/Toxicokinetics |
Z-Ala-Ala-Asn-AMC is generally considered to have low toxicity, consistent with its use as a research reagent. It is a peptide substrate with a coumarin fluorophore. Standard laboratory safety precautions, including the use of personal protective equipment, are recommended. No significant systemic toxicity is anticipated at typical research doses.
|
| References | |
| Additional Infomation |
Z-Ala-Ala-Asn-AMC (Cbz-Ala-Ala-Asn-AMC, CAS 149697-16-5) is a fluorogenic peptide substrate used for the enzyme legumain (EC 3.4.22.34). Its molecular formula is C₂₈H₃₁N₅O₈ and its molecular weight is 565.57 g/mol. It appears as a white powder. It is used as a standard substrate for legumain and is cleaved by legumain to release the fluorescent AMC group. It is intended for research use only.
|
| Molecular Formula |
C28H31N5O8
|
|---|---|
| Molecular Weight |
565.5744
|
| Exact Mass |
565.217
|
| CAS # |
149697-16-5
|
| PubChem CID |
90470169
|
| Appearance |
White to off-white solid powder
|
| LogP |
3.165
|
| Hydrogen Bond Donor Count |
5
|
| Hydrogen Bond Acceptor Count |
8
|
| Rotatable Bond Count |
12
|
| Heavy Atom Count |
41
|
| Complexity |
1040
|
| Defined Atom Stereocenter Count |
3
|
| SMILES |
CC1=CC(=O)OC2=C1C=CC(=C2)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)OCC3=CC=CC=C3
|
| InChi Key |
WVJTXYIHBUISHX-FIKGOQFSSA-N
|
| InChi Code |
InChI=1S/C28H31N5O8/c1-15-11-24(35)41-22-12-19(9-10-20(15)22)32-27(38)21(13-23(29)34)33-26(37)16(2)30-25(36)17(3)31-28(39)40-14-18-7-5-4-6-8-18/h4-12,16-17,21H,13-14H2,1-3H3,(H2,29,34)(H,30,36)(H,31,39)(H,32,38)(H,33,37)/t16-,17-,21-/m0/s1
|
| Chemical Name |
benzyl N-[(2S)-1-[[(2S)-1-[[(2S)-4-amino-1-[(4-methyl-2-oxochromen-7-yl)amino]-1,4-dioxobutan-2-yl]amino]-1-oxopropan-2-yl]amino]-1-oxopropan-2-yl]carbamate
|
| 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, 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)
|
| Solubility (In Vitro) |
DMSO : ~50 mg/mL (~88.41 mM)
H2O : < 0.1 mg/mL |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.42 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 (4.42 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 (4.42 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7681 mL | 8.8406 mL | 17.6813 mL | |
| 5 mM | 0.3536 mL | 1.7681 mL | 3.5363 mL | |
| 10 mM | 0.1768 mL | 0.8841 mL | 1.7681 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
