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题名: Microplate based assay for thrombin detection using an RNA aptamer as affinity ligand and cleavage of a chromogenic or a fluorogenic peptide substrate
作者: Hao, Lihua; Zhao, Qiang
刊名: MICROCHIMICA ACTA
出版日期: 2016-06
卷号: 183, 期号:6, 页码:1891-1898
关键词: Photometry ; Fluorescence ; Toggle-25 ; Enzyme substrate ; 2 '-deoxyfluoro sugars ; Microtiter plate ; Kinetic assay ; DNA aptamer ; Nuclease resistance
DOI: 10.1007/s00604-016-1833-4
部门归属: 环境化学与生态毒理学国家重点实验室
英文摘要: The authors report on a microplate based thrombin assay by using an RNA aptamer (Toggle-25) as the affinity ligand. The aptamer is nuclease-resistant because it contains nucleobases modified with 2'-deoxyfluoro saccharides. The aptamer is coated on the surface of the wells of microplates where it captures thrombin from a sample. Following a washing step, a chromogenic or a fluorogenic peptide substrate is added which is cleaved by the captured thrombin to form a yellow product (with absorbance at 405 nm) or a blue fluorescent product (with an emission maximum at 444 nm) after incubation at 37 A degrees C for 2 h. The assay enables thrombin to be determined with a 10 fM detection limit when using the fluorogenic substrate. The assay is selective in that several other proteins do not interfere. The assay can be applied to the determination of thrombin in diluted human serum and plasma.
收录类别: SCI
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内容类型: 期刊论文
URI标识: http://ir.rcees.ac.cn/handle/311016/35894
Appears in Collections:环境化学与生态毒理学国家重点实验室_期刊论文

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Recommended Citation:
Hao, Lihua,Zhao, Qiang. Microplate based assay for thrombin detection using an RNA aptamer as affinity ligand and cleavage of a chromogenic or a fluorogenic peptide substrate[J]. MICROCHIMICA ACTA,2016,183(6):1891-1898.
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文件名: Microplate based assay for thrombin detection using an RNA aptamer as affinity ligand and cleavage of a chromogenic or a fluorogenic peptide substrate.pdf
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