Suc-Leu-Tyr-AMC (Synonyms: SucLYAMC) |
| Katalog-Nr.GC44963 |
Suc-Leu-Tyr-AMC, a fluorescent substrate for calpain I and II and papain (cysteine protease) measuring the chymotrypsin-like peptidase activity of the 20S proteasome, emits bright fluorescence has been used extensively for calpain I and II and papain evaluation (Ex/Em: 360/460nm) .
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 94367-20-1
Sample solution is provided at 25 µL, 10mM.
Suc-Leu-Tyr-AMC, a fluorescent substrate for calpain I and II and papain (cysteine protease) measuring the chymotrypsin-like peptidase activity of the 20S proteasome, emits bright fluorescence has been used extensively for calpain I and II and papain evaluation (Ex/Em: 360/460nm) . Suc-Leu-Tyr-AMC causes a complete inhibition of casein breakdown at high concentrations that are regulated by Na+ and K+ in vivo[1,2]. Suc-Leu-Tyr-AMC is widely applied in calpain activity analysis in vitro[3]. Suc-Leu-Tyr-AMC can not be applied in live cells or in vivo, only extracellular calpain assay (in vitro or ex vivo) was applied.
Suc-Leu-Tyr-AMC is used to evaluate calpain activity with rat liver IMS protein (20μM; 20min incubation; 25°C)[4], fibrillating human atria lysate (125μM; 10min incubation; 37°C)[5], Mouse C2C12 myoblasts lysate (40mM; 30min; room temperature)[6] with extracted materials such as extracted proein and cell or tissue lysate.
References:
[1] Woo, K M et al. “Protease Ti from Escherichia coli requires ATP hydrolysis for protein breakdown but not for hydrolysis of small peptides.” *The Journal of biological chemistry* vol. 264,4 (1989): 2088-91.
[2] Seol, J H et al. “Na+, K+-specific inhibition of protein and peptide hydrolyses by proteasomes from human hepatoma tissues.” *FEBS letters* vol. 247,2 (1989): 197-200. doi:10.1016/0014-5793(89)81333-x
[3] Ozaki, Taku et al. “Characteristics of mitochondrial calpains.” *Journal of biochemistry* vol. 142,3 (2007): 365-76. doi:10.1093/jb/mvm143
[4] Ozaki, Taku et al. “Intravitreal injection or topical eye-drop application of a μ-calpain C2L domain peptide protects against photoreceptor cell death in Royal College of Surgeons' rats, a model of retinitis pigmentosa.” *Biochimica et biophysica acta*vol. 1822,11 (2012): 1783-95. doi:10.1016/j.bbadis.2012.07.018
[5] Goette, Andreas et al. “Calpains and cytokines in fibrillating human atria.” *American journal of physiology. Heart and circulatory physiology* vol. 283,1 (2002): H264-72. doi:10.1152/ajpheart.00505.2001
Liao, Zhiyin et al. “CHRNA1 induces sarcopenia through neuromuscular synaptic elimination.” *Experimental gerontology* vol. 166 (2022): 111891. doi:10.1016/j.exger.2022.111891
Protocal for in vitro calpain assay with fluorescence microplate reader[1]:
1.prepare sample (purified calpain‐1 from human or porcine erythrocytes in this case).
2.prepare Suc-Leu-Tyr-AMC (0.5mM) in 60 mM imidazole‐HCl buffer (5mM CaCl2, 5 mM cysteine, 2.5mM β‐mercaptoethanol, pH=7.3).
3.intiate reaction by adding the enzyme (purified calpain‐1 in this case) in Suc-Leu-Tyr-AMC solution, continued at 30°C for 15min, while the fluorescence of 7‐amino‐4‐methylcoumarin (Ex 380nm/Em 450nm) was monitored every 30s in a POLARstar Omega fluorescence microplate reader.
4.The rate of hydrolysis (increase in fluorescence/min) was determined from the linear portion of the curve.
5.The IC50 values were obtained by adjusting data from each experiment into a sigmoidal dose–response curve. The K i values were calculated from the average of the IC50 values and from a single substrate concentration by using a K i calculator tool for fluorescence‐based competitive binding assays (http://sw16.im.med.umich.edu/software/calc_ki/). The Km values of Suc‐Leu‐Tyr‐AMC used for the Ki calculation were 4.74 and 2.21mM for calpain‐1 and calpain‐2, respectively.
*This protocol only provides a guideline, and should be modified according to your specific needs
References:
[1]Baudry, Michel et al. “Identification and neuroprotective properties of NA-184, a calpain-2 inhibitor.” Pharmacology research & perspectives vol. 12,2 (2024): e1181. doi:10.1002/prp2.1181
| Cas No. | 94367-20-1 | SDF | |
| Überlieferungen | SucLYAMC | ||
| Canonical SMILES | CC(C1=CC=C(C=C1O2)NC([C@H](CC3=CC=C(C=C3)O)NC([C@@H](NC(CCC(O)=O)=O)CC(C)C)=O)=O)=CC2=O | ||
| Formula | C29H33N3O8 | M.Wt | 551.6 |
| Löslichkeit | 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 |
| 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 | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.8129 mL | 9.0645 mL | 18.1291 mL |
| 5 mM | 0.3626 mL | 1.8129 mL | 3.6258 mL |
| 10 mM | 0.1813 mL | 0.9065 mL | 1.8129 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
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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 14 reference(s) in Google Scholar.)
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