Suc-Leu-Leu-Val-Tyr-AMC (Synonyms: Suc-LLVY-AMC) |
Catalog No.GC44962
One-Click Copy Product Info
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Suc-Leu-Leu-Val-Tyr-AMC is a tetrapeptide compound that serves as a fluorogenic substrate for the 20S proteasome , chymotrypsin-like proteases , and calpains , with maximum excitation and emission wavelengths of 345 nm and 445 nm, respectively.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 94367-21-2
Sample solution is provided at 25 µL, 10mM.
Suc-Leu-Leu-Val-Tyr-AMC is a tetrapeptide compound that serves as a fluorogenic substrate for the 20S proteasome [1], chymotrypsin-like proteases [2], and calpains [3], with maximum excitation and emission wavelengths of 345 nm and 445 nm, respectively. Proteases such as calpains and 20S proteasome specifically recognize and cleave the "Leu-Leu-Val-Tyr" sequence, releasing the AMC fluorophore [4]，this makes it a valuable tool for studying calpain activity and proteasomal degradation.
References:
[1]. Stein RL, Melandri F, Dick L. Kinetic characterization of the chymotryptic activity of the 20S proteasome. Biochemistry. 1996 Apr 2;35(13):3899-908.
[2]. Gardner RC, Assinder SJ, Christie G, et al. Characterization of peptidyl boronic acid inhibitors of mammalian 20 S and 26 S proteasomes and their inhibition of proteasomes in cultured cells. Biochem J. 2000 Mar 1;346 Pt 2(Pt 2):447-54.
[3]. Debiasi RL, Squier MK, Pike B, et al. Reovirus-induced apoptosis is preceded by increased cellular calpain activity and is blocked by calpain inhibitors. J Virol. 1999 Jan;73(1):695-701. doi: 10.1128/JVI.73.1.695-701.1999.
[4]. Sasaki, T., T. Kikuchi, N. Yumoto, et al. 1984.Comparative specificity and kinetic studies on porcine calpain I and calpainII with naturally occurring peptides and synthetic fluorogenic substrates.J. Biol. Chem. 259:12489–12494.
Experimental Protocol for Suc-Leu-Leu-Val-Tyr-AMC
1.Preparation of Stock Solution: Dissolve Suc-Leu-Leu-Val-Tyr-AMC in DMSO to prepare a stock solution with a concentration of 1-10 mM. Store the stock solution at -20°C in the dark.
2.Prepare 1X Reaction Buffer: Prepare a reaction buffer containing 25 mM HEPES (pH 7.5), 10 mM DTT, 0.1% CHAPS, 0.5 mM PMSF, 2 μl/ml aprotinin.
3.Preparation of Working Solution: Dilute the Suc-Leu-Leu-Val-Tyr-AMC from the stock solution to 20 μM.
4.Preparation of Sample: Prepare sample lysates containing the proteasomal to be assayed or purified 20S and 26S proteasomes.
5.Add 50 μl of cytosolic proteasomal (containing 40-75 μg protein) to 0.5 ml of working solution, incubate at 37°C for 30 minutes.
6.Fluorescence Measurement: After incubation at 37°C for 30 minutes, measure the fluorescence with a fluorometer, using excitation and emission wavelengths of 345 nm and 445 nm, respectively.
This protocol only provides a guideline and should be modified according to your specific needs.
References:
[1]. Tadlock L, Yamagiwa Y, Hawker J, Marienfeld C, Patel T. Transforming growth factor-beta inhibition of proteasomal activity: a potential mechanism of growth arrest. Am J Physiol Cell Physiol. 2003 Aug;285(2):C277-85. doi: 10.1152/ajpcell.00550.2002. Epub 2003 Mar 19. PMID: 12646415.
| Cas No. | 94367-21-2 | SDF | |
| Synonyms | Suc-LLVY-AMC | ||
| Canonical SMILES | CC(C1=CC=C(C=C1O2)NC([C@H](CC3=CC=C(C=C3)O)NC([C@@H](NC([C@@H](NC([C@@H](NC(CCC(O)=O)=O)CC(C)C)=O)CC(C)C)=O)C(C)C)=O)=O)=CC2=O | ||
| Formula | C40H53N5O10 | M.Wt | 763.9 |
| Solubility | DMF: 30 mg/ml,DMSO: 30 mg/ml,DMSO:PBS(pH7.2) (1:1): 0.5 mg/ml,Ethanol: 20 mg/ml | Storage | Store at -20°C, protect from light, stored under nitrogen |
| 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 | 1.3091 mL | 6.5454 mL | 13.0907 mL |
| 5 mM | 261.8 μL | 1.3091 mL | 2.6181 mL |
| 10 mM | 130.9 μL | 654.5 μL | 1.3091 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 14 reference(s) in Google Scholar.)