SensoLyte® 490 HCV Protease Assay Kit *Fluorimetric*
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产品英文名称:SensoLyte® 490 HCV Protease Assay Kit *Fluorimetric*
产品中文名称:SensoLyte® 490 HCV 蛋白酶荧光检测试剂盒
SensoLyte® 490 HCV Protease Assay Kit *Fluorimetric* 产品介绍
丙型肝炎病毒(HCV)NS3/4A蛋白酶是病毒在NS3-NS4A,ns4a-ns4b,ns4b-ns5a,和ns5a-ns5b这些位点非结构蛋白裂解所需的。这些裂解的病毒蛋白的成熟至关重要。因此,该蛋白酶成为开发抗HCV药物的关键目标之一。该 SensoLyte®490 HCV蛋白酶检测试剂盒提供了方便的测定。HCV NS3/4A蛋白酶抑制剂和连续定量高通量筛选HCV NS3/4A蛋白酶活性采用5-FAM / QXL™490.荧光共振能量转移(FRET)肽。这个肽序列是来自NS4A /细胞裂解位点的半胱氨酸。自然裂解位点与γ-氨基丁酸取代(阿布)和易断裂的肽键以酯键。这些修改提高kcat / Km值的100倍以上,使检测的活性的NS3/4A在纳摩尔浓度的蛋白酶。在FRET肽,荧光淬灭的EDANS DABCYL。在分裂成两个单独的片段的HCV NS3/4A蛋白酶在阿布阿拉债券(见方案1),在荧光EDANS恢复,可以监测在340 nm的激发/发射= 490n实验是在一个方便的96孔或384孔微孔板格式进行。
SensoLyte® 490 HCV 蛋白酶荧光检测试剂盒的成分
Component A HCV NS3/4A protease substrate 250 μL
EDANS/DABCYL FRET peptide, Ex/Em=340
nm/490 nm upon cleavage
Component B EDANS, fluorescence reference standard 100 μM DMSO
Ex/Em=340 nm/490 nm solution, 10 μL
Component C 2X Assay buffer 25 mL
Component D Stop solution 15 mL
Component E DTT 1 M,1 mL
Component F Pep4AK HCV NS3 protease cofactor 150 μL, 600 μM
SensoLyte® 490 HCV Protease Assay Kit *Fluorimetric* 的保存
• Store all kit components at -20°C
• Protect Components A and B from light
• Components C and D can be stored at room temperature for convenience
SensoLyte® 490 HCV 蛋白酶荧光检测试剂盒的英文描述
HCV protease is identified as an important drug-screening target. The SensoLyte® 490 HCV Protease Assay Kit uses an optimized FRET peptide substrate for the continuous measurement of HCV NS3/4a activity. This FRET substrate is cleaved specifically by HCV NS3/4a protease thereby liberating the C-terminal peptide-fluorophore fragment from the proximity quenching effect of the dark quencher, resulting in a more than 10-fold increase in fluorescence. This assay can detect HCV NS3/4a protease activity at 1 pmole. The kit contains: • EDANS/DABCYL-based FRET peptide substrate (Ex/Em=340/490 nm upon cleavage) • Assay buffer • Fluorescence reference standard for calibration • A detailed protocol.
SensoLyte® 490 HCV Protease Assay Kit *Fluorimetric* 系列产品订购信息
AnaSpec 61017-5 HCV NS3/4A protease genotype 1b, recombinant 5 µg
AnaSpec 61017-10 HCV NS3/4A protease genotype 1b, recombinant 10 µg
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