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COVID-19肽底物Covidyte TF670

英文名称:Covidyte™ TF670
COVID-19肽底物Covidyte TF670
价格 4042
产品规格
100 tests

产品货号
产品参数
Ex (nm)649Em (nm)664
分子量3620.33溶剂DMSO
存储条件在零下15度以下保存, 避免光照
产品概述

冠状病毒(CoV)是一种可以感染人类和多种动物,因其形态看上去像中世纪欧洲帝的皇冠,因此命名为“冠状病毒”。这类病毒具有胃肠道、呼吸道和神经系统的嗜性,可以导致多种疾病。在2019年底,一种新型的冠状病毒被称为严重急性呼吸系统综合症冠状病毒2(SARS-CoV-2),开始在全球蔓延。即使大多数受感染的患者仅患有轻度症状,例如发烧和咳嗽,但该疾病可能发展为致命的肺炎和急性呼吸衰竭病例,尤其是在多病体弱的的老人中。该病毒仅仅从出现开始的三四个月,全球已经感染了超过100万人。当前,尚无用于Covid-19的任何具体有效的选择。目前,Covid-19的临床主要是对症,并结合上市的抗病毒(如瑞德昔韦和)以继发感染。目前全都迫切需要开发抗SARS-CoV-2的特异性抗病毒方法和疫苗,而冠状病毒主蛋白酶是抗CoV设计的重要靶点,在病毒基因表达和复制中起着关键作用。目前已掌握的人类免疫缺陷病毒(HIV)和丙型肝炎的成功策略是抑制蛋白水解酶消灭病毒蛋白酶。而且证明了蛋白酶抑制剂病毒感染具有非常大的潜力。同样,SARS-CoV-2病毒必须要进行复制的酶便是蛋白酶,因此Covid-19是抗病毒极好的靶点。

Covidyte TF670是一种包含11个氨基酸序列的肽底物,可被冠状病毒蛋白酶切割。FRET肽的"N"和"C"末端分别包含Dabcyl(淬灭剂)和Edans(供体),当该肽完整时,Edans的荧光可被Dabcy有效地淬灭。当该肽被冠状病毒蛋白酶水解时,Edans片段产生明显增强的荧光,因为其荧光不再被Dabcyl淬灭。冠状病毒蛋白酶的活性可以通过Edans的荧光强度有效地检测到。Covidyte TF670是筛选冠状病毒蛋白酶抑制剂的便捷工具。百萤生物是AAT Bioquest的中国代理商,为您提供优质的Covidyte肽底物系列产品。 

点击查看光谱

 

适用仪器


荧光酶标仪  
Ex: 640nm
Em: 680nm
Cutoff: 660nm
推荐孔板: 纯黑色孔板
实验方案

样品实验方案

溶液配制

储备溶液配制

1.Covidyte TF670储备液(200X):向Covidyte TF670小瓶中添加25 µL(对于cat#13540)或250 µL(对于cat#13541)DMSO。

注意:制成单次使用的等分试样,并储存在-20°C下。

 

工作溶液配制

1.Covidyte TF670工作溶液在20 mM Tris缓冲液(pH 7.5)或自备缓冲液中以1:200稀释底物储备液。每个实验在96孔板中使用50μL底物溶液。

2.冠状病毒蛋白酶稀释:根据需要稀释冠状病毒蛋白酶。

 

实验步骤

(一个96孔板的样品方案)

  1. 将50μLCovidyte TF670工作溶液和50μL冠状病毒蛋白酶稀释液添加到测定板的所有孔中。
  2. 使用荧光酶标仪在Ex / Em = 640/680 nm(截止660nm)处检测荧光的增加。 

对于动力学读数:立即开始连续不断地测量荧光强度,并每5分钟记录一次数据,持续30-120分钟。
对于终点读数:在避光的条件下,将反应在所需温度下孵育30至120分钟。然后测量荧光强度。
 

图示

 

图1蛋白酶在蛋白质、细胞调节和信号转导以及氨基酸的生成中起着至关重要的作用,用于蛋白质合成或其他代谢途径。 FRET蛋白酶底物广泛用于检测蛋白酶活性,特别是用于病毒蛋白酶的检测,这些病毒蛋白酶经常需要较长的肽序列才能实现佳结合,例如冠状病毒,HIV和HCV蛋白酶。原理:内部淬灭的FRET肽底物被蛋白酶消化,荧光强度与蛋白酶活性成正比,从而产生高荧光的肽片段。Tide QuencherTide Fluor 和iFluor 染料都是研发用于高通量筛选应用的FRET蛋白酶底物的极其有效的淬灭剂

 

 

相关产品

品牌 品名 货号 规格 描述
AAT Covidyte IF670 #13542 100 Tests

1.包含可被冠状病毒蛋白酶切割的14个氨基酸序列的肽底物

2.#13542:当该肽被冠状病毒蛋白酶水解时,iFluor 670片段产生明显增强的荧光,因为其荧光不再被TQ5淬灭;#13541:当该肽被冠状病毒蛋白酶水解时,TF5片段产生明显增强的荧光,因为其荧光不再被TQ5淬灭

AAT Covidyte TF670 #13541 1000 Tests
AAT Covidyte EN450 #13535 100 Tests
AAT Covidyte ED450 #13537 100 Tests 包含可被冠状病毒蛋白酶切割的11个氨基酸序列的肽底物

 

参考文献

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Learning from the Past: Possible Urgent Prevention and Treatment Options for Severe Acute Respiratory Infections Caused by 2019-nCoV.
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SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor.
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Journal: Cell (2020)

Structural Basis for Inhibiting Porcine Epidemic Diarrhea Virus Replication with the 3C-Like Protease Inhibitor GC376.
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α-Ketoamides as Broad-Spectrum Inhibitors of Coronavirus and Enterovirus Replication: Structure-Based Design, Synthesis, and Activity Assessment.
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Characterization of amino acid substitutions in feline coronavirus 3C-like protease from a cat with feline infectious peritonitis treated with a protease inhibitor.
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Evaluation of a non-prime site substituent and warheads combined with a decahydroisoquinolin scaffold as a SARS 3CL protease inhibitor.
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Efficacy of a 3C-like protease inhibitor in treating various forms of acquired feline infectious peritonitis.
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Protease inhibitors broadly effective against feline, ferret and mink coronaviruses.
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