tailieunhanh - Báo cáo khoa học: Selective detection of superoxide anion radicals generated from macrophages by using a novel fluorescent probe

Quantitation of superoxide radical (O2 – Æ) production at the site of radical generation remains challenging. A simple method to detect nanomolar to micromolar levels of superoxide radical in aqueous solution has been devel-oped and optimized. This method is based on the efficient trapping of O2 – Æ using a novel fluorescent probe (2-chloro-1,3-dibenzothiazolinecyclohex-ene), coupled with a spectra character-signaling increase event. | ỊFEBS Journal Selective detection of superoxide anion radicals generated from macrophages by using a novel fluorescent probe Jing Jing Gao1 Ke Hua Xu1 Bo Tang1 Ling Ling Yin1 Gui Wen Yang2 and Li Guo An2 1 College of Chemistry ChemicalEngineering and Materials Science Shandong Normal university Jinan China 2 College of Life Science Shandong NormalUniversity Jinan China Keywords 2-chloro-1 3-dibenzothiazolinecyclohexene fluorescence image fluorescent probe macrophages superoxide anion radical Correspondence B. Tang College of Chemistry Chemical Engineering and Materials Science Engineering Research Center of Pesticide and Medicine Intermediate Clean Production Ministry of Education Shandong NormalUniversity Jinan 250014 China Fax 86 531 861 80017 Tel 86 531 861 80010 E-mail tangb@ Received 24 October 2006 revised 11 January 2007 accepted 29 January 2007 doi Quantitation of superoxide radical O 2 production at the site of radical generation remains challenging. A simple method to detect nanomolar to micromolar levels of superoxide radical in aqueous solution has been developed and optimized. This method is based on the efficient trapping of O2- using a novel fluorescent probe 2-chloro-1 3-dibenzothiazolinecyclohex-ene coupled with a spectra character-signaling increase event. A high-specificity and high-sensitivity fluorescent probe was synthesized in-house and used to image O2- in living cells. Better selectivity for O2 over competing cellular reactive oxygen species and some biological compounds illustrates the advantages of our method. Under optimal conditions the linear calibration range for superoxide anion radicals was X X 10-6 M. The detection limit was X 10-9 M. Fluorescence images of probe-stained macrophages stimulated with 4b-phorbol 12-myristate 13-acetate were obtained successfully using a confocal laser scanning microscope. Reactive oxygen species ROS such as the superoxide anion radical .

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