Bz-Nle-Lys-Arg-Arg-AMC (Synonyms: Benzoyl-Nle-Lys-Arg-Arg-AMC, Benzoyl-Nle-Lys-Arg-Arg-7-amino-4-methylcoumarin, Bz-Nle-Lys-Arg-Arg-AMC) |
| Catalog No.GA21221 |
Bz-Nle-Lys-Arg-Arg-AMC is a fluorogenic tetra-peptide substrate for yellow fever virus (YFV) non-structural 3 (NS3), dengue virus (DV) NS2B/3 serine proteases, and Zika virus (ZIKV) NS2B/NS3 serine proteases.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 863975-32-0
Sample solution is provided at 25 µL, 10mM.
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Bz-Nle-Lys-Arg-Arg-AMC is a fluorogenic tetra-peptide substrate for yellow fever virus (YFV) non-structural 3 (NS3), dengue virus (DV) NS2B/3 serine proteases, and Zika virus (ZIKV) NS2B/NS3 serine proteases [1,2,3].
After enzymatic hydrolysis by YNS2B/NS3 serine proteases, Bz-Nle-Lys-Arg-Arg-AMC releases 7-amino-4-methylcoumarin (AMC), whose fluorescence can be used to determine the activity of YNS2B/NS3 serine proteases. AMC exhibits excitation/emission maxima at 340-360/440-460 nm, respectively.
References:
[1]. Ulanday GE, Okamoto K, Morita K. Development and utility of an in vitro, fluorescence-based assay for the discovery of novel compounds against dengue 2 viral protease. Tropical Medicine and Health. 2016 Dec;44:1-0.
[2]. Loehr K, Knox JE, Phong WY, Ma NL, Yin Z, Sampath A, Patel SJ, Wang WL, Chan WL, Rao KR, Wang G. Yellow fever virus NS3 protease: peptide-inhibition studies. Journal of general virology. 2007 Aug;88(8):2223-7.
[3]. Lee H, Ren J, Nocadello S, Rice AJ, Ojeda I, Light S, Minasov G, Vargas J, Nagarathnam D, Anderson WF, Johnson ME. Identification of novel small molecule inhibitors against NS2B/NS3 serine protease from Zika virus. Antiviral research. 2017 Mar 1;139:49-58.
| Cell experiment [1]: | |
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Preparation Method |
FRET assay was performed to evaluate NS2B-NS3 protease activity and its inhibition by flavonoids. NS2B-NS3 protease in buffer solution (20 mM HEPES, 20 % glycerol and 1 mM CHAPS, pH 9.0) was pre-incubated at 37 °C for 30 min, and the reaction was initiated by adding 100 μM of the substrate Bz-Nle-Lys-Arg-Arg-AMC. The fluorescence was monitored per 15 s in 10 min with a microplate spectrophotometer at 360 nm and 450 nm for excitation and emission, respectively. The inhibition assay was performed in 100 μL of buffer containing 20 mM HEPES (pH 9.0), 20 % glycerol, 1 mM CHAPS, 0.1 μM NS2B-NS3 protease and 100 μM substrate with varying concentrations of inhibitors. NS2B-NS3 protease was pre-incubated with inhibitors at 37°C for 30 min, and the reaction was initiated by adding 100 μM of the substrate Bz-Nle-Lys-ArgArg-AMC. After 10 min, the fluorescence was monitored with a microplate spectrophotometer at 360 nm and 450 nm for excitation and emission, respectively. Aprotinin (1 μM) was used as a positive control. |
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Reaction Conditions |
100 µM for 10 min |
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References: [1]: Zou M, Liu H, Li J, Yao X, Chen Y, Ke C, Liu S. Structure-activity relationship of flavonoid bifunctional inhibitors against Zika virus infection. Biochemical pharmacology. 2020 Jul 1;177:113962. |
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| Cas No. | 863975-32-0 | SDF | |
| Synonyms | Benzoyl-Nle-Lys-Arg-Arg-AMC, Benzoyl-Nle-Lys-Arg-Arg-7-amino-4-methylcoumarin, Bz-Nle-Lys-Arg-Arg-AMC | ||
| Formula | C41H60N12O7 | M.Wt | 833 |
| Solubility | DMF: 30mg/mL,DMSO: 30mg/mL,DMSO:PBS (pH 7.2) (1:1): 0.5mg/mL,Ethanol: 20mg/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. |
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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 | 1.2005 mL | 6.0024 mL | 12.0048 mL |
| 5 mM | 0.2401 mL | 1.2005 mL | 2.401 mL |
| 10 mM | 0.12 mL | 0.6002 mL | 1.2005 mL |
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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.
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