Z-Leu-Arg-AMC |
Katalog-Nr.GA23847
One-Click Copy Product Info
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Z-Leu-Arg-AMC is a 4-methylcoumarinyl-7-amide (AMC) leucine derivative with carboxybenzoyl (Z).
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 156192-32-4
Sample solution is provided at 25 µL, 10mM.
Z-Leu-Arg-AMC is a 4-methylcoumarinyl-7-amide (AMC) leucine derivative with carboxybenzoyl (Z). Z-Leu-Arg-AMC can be cleaved by cathepsin B, L, and S, and is mainly used to detect the enzyme activity of cysteine proteases such as cathepsin B[1][2][3].
References:
[1] Yoon MC, Phan V, Podvin S, Mosier C, O'Donoghue AJ, Hook V. Distinct Cleavage Properties of Cathepsin B Compared to Cysteine Cathepsins Enable the Design and Validation of a Specific Substrate for Cathepsin B over a Broad pH Range. Biochemistry. 2023;62(15):2289-2300.
[2] Song Y, Fan H, Anderson MJ, et al. Electrochemical Activity Assay for Protease Analysis Using Carbon Nanofiber Nanoelectrode Arrays. Anal Chem. 2019;91(6):3971-3979.
[3]BRENDA Enzyme Database. Cathepsin L (EC 3.4.22.15) substrate information. Technische Universität Braunschweig. Accessed March 10, 2026. https://www.brenda-enzymes.org
Cathepsin B Activity Assay[1-2]
This plan is based on literature, please modify it to meet your specific needs.
1. Preparation of substrate working solution
(1) Prepare storage solution: Dissolve Z-Leu-Arg-AMC in DMSO.
Note: Unused storage solution should be aliquoted and stored in the dark at -20°C or -80°C to avoid repeated freezing and thawing.
(2) Prepare working solution: Dilute the stock solution with MES buffer (containing 5mM dithiothreitol (DTT) and 25mM 2-(4-Morpholino) ethanesulfonic acid, pH=5.0).
Note: Prepare the working solution fresh and store briefly at 4°C or on ice.
2. Pre-incubate in MES buffer at 37°C to activate cathepsin B, typically at a concentration of 0.1-1μg/mL.
3. Prepare a 96-well plate, including:
Sample blank wells: Z-Leu-Arg-AMC substrate only, concentration 40-100μM.
Substrate blank wells: activated cathepsin B only, concentration 0.1-1μg/mL.
Experimental wells: a mixture of Z-Leu-Arg-AMC substrate and cathepsin B, concentrations of 0.1-1μg/mL and 40-100μM, respectively.
4. Incubate the 96-well plate at 37℃ for 10-60 min, and detect the fluorescence intensity at 440-460nm under 350-360nm excitation using a microplate reader.
Precautions:
1) Fluorescent substrate has quenching problems. Please avoid light as much as possible to slow down fluorescence quenching.
2) For your safety and health, please wear a lab coat and disposable gloves.
References:
[1] Yoon MC, Phan V, Podvin S, Mosier C, O'Donoghue AJ, Hook V. Distinct Cleavage Properties of Cathepsin B Compared to Cysteine Cathepsins Enable the Design and Validation of a Specific Substrate for Cathepsin B over a Broad pH Range. Biochemistry. 2023;62(15):2289-2300.
[2] Song Y, Fan H, Anderson MJ, et al. Electrochemical Activity Assay for Protease Analysis Using Carbon Nanofiber Nanoelectrode Arrays. Anal Chem. 2019;91(6):3971-3979.
| Cas No. | 156192-32-4 | SDF | |
| Formula | C30H38N6O6 | M.Wt | 578.67 |
| Löslichkeit | DMSO : 100 mg/mL (172.81 mM; Need ultrasonic) | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.7281 mL | 8.6405 mL | 17.281 mL |
| 5 mM | 345.6 μL | 1.7281 mL | 3.4562 mL |
| 10 mM | 172.8 μL | 864.1 μL | 1.7281 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 25 reference(s) in Google Scholar.)