SensoLyte? 520 HCV Protease Assay Kit *Fluorimetric*
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產(chǎn)品英文名稱:SensoLyte® 520 HCV Protease Assay Kit *Fluorimetric*
產(chǎn)品中文名稱:SensoLyte® 520 HCV 蛋白酶熒光檢測試劑盒
SensoLyte® 520 HCV Protease Assay Kit *Fluorimetric* 產(chǎn)品介紹
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這些位點非結(jié)構(gòu)蛋白裂解所需的。這些裂解的病毒蛋白的成熟至關重要。因此,該蛋白酶成為開發(fā)抗HCV藥物的關鍵目標之一。該 SensoLyte®520 HCV蛋白酶檢測試劑盒提供了方便的測定。HCV NS3/4A蛋白酶抑制劑和連續(xù)定量高通量篩選HCV NS3/4A蛋白酶活性采用5-FAM / QXL™520.熒光共振能量轉(zhuǎn)移(FRET)肽。這個肽序列是來自裂解位點NS4A / NS4B。在這個肽中,5-FAM熒光淬滅由QXL™520完成。在分裂成兩個獨立的片段的HCV NS3/4A蛋白酶(方案1),在熒光5-FAM回收,可在激發(fā)/發(fā)射= 490 nm / 520nm被監(jiān)測。具有優(yōu)異的熒光量子產(chǎn)率和更長的波長,5-FAM信號不受細胞成分和測試化合物的熒光。測定檢測到低至1.56毫微克/毫升活躍的HCV NS3/4A蛋白酶。
SensoLyte® 520 HCV蛋白酶熒光檢測試劑盒的成分
Component A HCV NS3/4A protease substrate 120 μL
Ex/Em=490 nm/520 nm upon cleavage
Component B 5-FAM, fluorescence reference standard 100 μM, 5 μL
Ex/Em=490 nm/520 nm upon cleavage
Component C 2X Assay buffer 10 mL
Component D Stop solution 10 mL
Component E DTT 1 M, 0.5 mL
Component F Pep4AK 50μL, 600μM
SensoLyte® 520 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® 520 HCV 蛋白酶熒光檢測試劑盒的英文描述
The SensoLyte® 520 HCV Assay Kit uses a new FRET peptide substrate that incorporates 5-FAM (fluorophore) and QXL™ 520 (quencher) for a continuous measurement of enzyme activities. In the intact FRET peptide, the fluorescence of 5-FAM is quenched by QXL™ 520. Upon cleavage of the FRET peptide by HCV NS3/4a protease, the fluorescence of 5-FAM is recovered and can be continuously monitored at excitation/emission = 490 nm/520 nm. With superior fluorescence quantum yield and longer emission wavelength, this 5-FAM/QXL™ 520-based FRET peptide shows less interference from autofluorescence of test compounds and cellular components. This assay is ten times more sensitive than an EDANS/DABCYL based assay and can detect 0.1 pmole of HCV NS3/4a protease. The assays are performed in a convenient 96-well or 384-well microplate format. The kit contains: • 5-FAM/QXL™ 520-based FRET peptide substrate (Ex/Em=490/520 nm upon cleavage) • Fluorescence reference standard for calibration • A detailed protocol.
SensoLyte® 520 HCV Protease Assay Kit *Fluorimetric* 系列產(chǎn)品訂購信息
AnaSpec 71145 SensoLyte® 520 HCV Protease Assay Kit *Fluorimetric* 1 kit
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