Z-Arg-Leu-Arg-Gly-Gly-AMC |
| رقم الكتالوجGA23715 |
Z-RLRGG-AMC هو مركب فلوروجيني لعامل الإزالة الدي يوبيكتينية isopeptidase T (IPaseT) وغيره من إنزيمات تحطم نهاية سلسلة الأحماض الأمينية في عائدات التشابك (UCHs) بناءً على أطراف C من ubiquitin ، وذلك باستخدام قيمة kcat / Km 95 M؟¹s؟¹. يستخدم بالتعاون مع Z-Leu-Arg-Gly-Gly-AMC لدينا في اختبارات مستمرة للدراسات الميكانيكية الدقيقة أو اختبارات صفائف المجاميل لغرض فحص المثبطات بشكل سريع.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 167698-69-3
Sample solution is provided at 25 µL, 10mM.
Z-Arg-Leu-Arg-Gly-Gly-AMC (Z-RLRGG-AMC) is a peptide substrate of SARS-CoV papain (PLpro), where RLRGG is the C-terminal sequence of ubiquitin (Ub)[1, 2]. The maximum excitation/emission wavelengths of Z-Arg-Leu-Arg-Gly-Gly-AMC are 340/450 nm, respectively [3].
Z-Arg-Leu-Arg-Gly-Gly-AMC is primarily used to study protease activity. By measuring the fluorescence intensity of released 7-amino-4-methylcoumarin (AMC), the catalytic activity of proteases can be assessed[4]. Z-Arg-Leu-Arg-Gly-Gly-AMC consists of a pentapeptide sequence linked to the fluorescent group AMC. The "Z" denotes the benzyloxycarbonyl (Z) protecting group, which is used to protect amino acids from nonspecific reactions [5].
References:
[1] Báez-Santos Y M, Mielech A M, Deng X, et al. Catalytic function and substrate specificity of the papain-like protease domain of nsp3 from the Middle East respiratory syndrome coronavirus[J]. Journal of virology, 2014, 88(21): 12511-12527.
[2] Li L, Ma L, Hu Y, et al. Natural biflavones are potent inhibitors against SARS-CoV-2 papain-like protease[J]. Phytochemistry, 2022, 193: 112984.
[3] Fu Z, Huang B, Tang J, et al. The complex structure of GRL0617 and SARS-CoV-2 PLpro reveals a hot spot for antiviral drug discovery[J]. Nature communications, 2021, 12(1): 488.
[4] Wang C, Chen S, Yeo S, et al. Elevated p62/SQSTM1 determines the fate of autophagy-deficient neural stem cells by increasing superoxide[J]. Journal of Cell Biology, 2016, 212(5): 545-560.
[5] Kadereit D, Waldmann H. Enzymatic protecting group techniques[J]. Chemical Reviews, 2001, 101(11): 3367-3396.
This plan only provides a guide, please modify it to meet your specific needs.
1. Solution Preparation:
(1) Stock Solution: Dissolve Z-Arg-Leu-Arg-Gly-Gly-AMC in DMSO to a final concentration of 1 mM.
Note: Aliquot the unused stock solution and store at -20°C in the dark. Avoid repeated freeze-thaw cycles.
(2) Working Solution: Dilute the stock solution with assay buffer to the desired working concentration.
Note: Adjust the optimal working concentration according to the actual conditions or refer to the literature for gradient concentration tests. The working solution must be prepared fresh for each use.
2. Steps for Measuring PLpro Activity Using Z-Arg-Leu-Arg-Gly-Gly-AMC[1] (from the literature, for reference only):
(1) Prepare the assay buffer containing 50 mM HEPES (pH 7.4), 0.01% Triton X-100 (v/v), 0.1 mg/mL BSA, and 2 mM DTT. Set up the reaction system in a 384-well plate, with a final reaction volume of 50 μL per well.
(2) Add PLpro to each well to a final concentration of 100 nM.
(3) Add 5 μL of Z-Arg-Leu-Arg-Gly-Gly-AMC stock solution to each well, achieving a final concentration of 50 μM.
(4) Substrate hydrolysis was monitored using a microplate reader. The excitation wavelength is 340 nm, and the emission wavelength is 450 nm. Record the fluorescence signal every 3 minutes.
Note: This protocol provides guidance for measuring PLpro activity using Z-Arg-Leu-Arg-Gly-Gly-AMC. Adjustments can be made according to other literature and specific experimental requirements.
References:
[1]Fu Z, Huang B, Tang J, et al. The complex structure of GRL0617 and SARS-CoV-2 PLpro reveals a hot spot for antiviral drug discovery[J]. Nature communications, 2021, 12(1): 488.
| Cas No. | 167698-69-3 | SDF | |
| Formula | C40H56N12O9 | M.Wt | 848.96 |
| الذوبان | Soluble in DMSO | 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. |
||
| 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 | 1.1779 mL | 5.8896 mL | 11.7791 mL |
| 5 mM | 0.2356 mL | 1.1779 mL | 2.3558 mL |
| 10 mM | 0.1178 mL | 0.589 mL | 1.1779 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 ddH2O, 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%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 4 reference(s) in Google Scholar.)
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