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) 目录号 : GA21221
Bz-Nle-Lys-Arg-Arg-AMC is a fluorogenic tetra-peptide substrate for yellow fever virus (YFV) non-structural 3 (NS3).
Cas No.:863975-32-0
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
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| 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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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.
Bz-Nle-Lys-Arg-Arg-AMC 是黄热病病毒 (YFV) 非结构 3 (NS3)、登革热病毒 (DV) NS2B/3 丝氨酸蛋白酶和寨卡病毒的荧光四肽底物(ZIKV) NS2B/NS3 丝氨酸蛋白酶 [1,2,3]。
在 YNS2B/NS3 丝氨酸蛋白酶酶促水解后，Bz-Nle-Lys-Arg-Arg-AMC 释放 7-amino-4 -甲基香豆素 (AMC)，其荧光可用于确定 YNS2B/NS3 丝氨酸蛋白酶的活性。 AMC 的激发/发射最大值分别位于 340-360/440-460 nm。
| Cas No. | 863975-32-0 | SDF | |
| 别名 | Benzoyl-Nle-Lys-Arg-Arg-AMC, Benzoyl-Nle-Lys-Arg-Arg-7-amino-4-methylcoumarin, Bz-Nle-Lys-Arg-Arg-AMC | ||
| 分子式 | C41H60N12O7 | 分子量 | 833 |
| 溶解度 | DMF: 30mg/mL,DMSO: 30mg/mL,DMSO:PBS (pH 7.2) (1:1): 0.5mg/mL,Ethanol: 20mg/mL | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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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 |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。