A label-free assay for high sensitive detection of RNase based on two near IR fluorescence probes
| Autor: | Wei Yang, Changying Yang, Qingyun Gao, Jinya Du, Zhouxuan Xiang, Yuzhi Dong, Na Huang |
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| Rok vydání: | 2018 |
| Předmět: |
Detection limit
biology Chemistry RNase P 010401 analytical chemistry Biophysics RNA Substrate (chemistry) General Chemistry 010402 general chemistry Condensed Matter Physics 01 natural sciences Biochemistry Fluorescence Atomic and Molecular Physics and Optics 0104 chemical sciences RNA silencing biology.protein Enzyme kinetics RNase H |
| Zdroj: | Journal of Luminescence. 204:162-168 |
| ISSN: | 0022-2313 |
| DOI: | 10.1016/j.jlumin.2018.07.048 |
| Popis: | RNase, whose function was cleaving RNA in ssRNA, dsRNA or DNA-RNA hybrid chain, can be analyzed directly by fluorescence probe assisted with RNA. In this paper, we constructed a none-labeled RNase assay based on fluorescence probe with high sensitivity and specificity. Two TICT characterize probes (H2 and L2) exhibited strong luminescence when bound with RNA. Then RNase hydrolysis substrate RNA exposing probe into buffer and resulted in fluorescence quench, causing “OFF-ON-OFF” fluorescence switch. We successfully applied the assay to detect two kind of RNase (RNase A and RNase H) with the detection limit of 1.67 × 10−5 U/mL for RNase A and 1.06 × 10−4 U/mL for RNase H, respectively. Enzyme kinetics confirmed again the specific cleaving process. Moreover, this approach could also be used for the screening of RNase inhibitors. RNaseOUT was active against RNase A with the IC50 values of 0.88 U/mL, but not against RNase H. |
| Databáze: | OpenAIRE |
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