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The assay is based on the indication of the enzymatic consumption of H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub> (HP), using a fluorescent europium\u2013tetracycline (Eu<jats:sub>3<\/jats:sub>TC) complex as indicator. On addition of HP, this complex forms a highly fluorescent adduct (Eu<jats:sub>3<\/jats:sub>TC\u2013HP), which is decomposed in the presence of POx to form the weakly fluorescent europium\u2013tetracycline (Eu<jats:sub>3<\/jats:sub>TC). Hence, the activity of the enzyme can be directly determined by means of the luminescent Eu<jats:sub>3<\/jats:sub>TC complex as indicator. The POx assay demonstrated herein was elaborated starting from a spectral characterization of the complex systems involved. Due to the long lifetime of lanthanide luminescence, both steady\u2010state and time\u2010resolved luminescence assays can easily be performed. The time\u2010resolved assay can quantify POx in the range from 4.0\u00d710<jats:sup>\u22125<\/jats:sup> to 5.9\u00d710<jats:sup>\u22123<\/jats:sup> U\u2009mL<jats:sup>\u22121<\/jats:sup>, with a limit of detection of 1.0\u00d710<jats:sup>\u22125<\/jats:sup> U\u2009mL<jats:sup>\u22121<\/jats:sup>. The effects of POx inhibitors such as cyanide, hydroxylamine, and azide have also been studied. In addition, a time\u2010resolved fluorescent detection method for a POx\u2010based enzyme\u2010linked immunosorbent assay (ELISA) has been developed, which is demonstrated in a sandwich model assay with bovine IgG serving as analyte. Furthermore, a time\u2010resolved fluorescent imaging method is demonstrated that makes use of a straightforward imaging set\u2010up adjusted to the optical properties of the europium reagent.<\/jats:p>","DOI":"10.1002\/chem.200500884","type":"journal-article","created":{"date-parts":[[2006,1,17]],"date-time":"2006-01-17T06:48:41Z","timestamp":1137480521000},"page":"2730-2738","source":"Crossref","is-referenced-by-count":28,"title":["A Novel Method for Time\u2010Resolved Fluorimetric Determination and Imaging of the Activity of Peroxidase, and Its Application to an Enzyme\u2010Linked Immunosorbent Assay"],"prefix":"10.1002","volume":"12","author":[{"given":"Zhihong","family":"Lin","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meng","family":"Wu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Otto S.","family":"Wolfbeis","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Sch\u00e4ferling","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2006,3,10]]},"reference":[{"unstructured":"\u00a0","key":"e_1_2_6_1_2"},{"doi-asserted-by":"publisher","key":"e_1_2_6_2_2","DOI":"10.1002\/ange.200454141"},{"doi-asserted-by":"publisher","key":"e_1_2_6_2_3","DOI":"10.1002\/anie.200454141"},{"doi-asserted-by":"publisher","key":"e_1_2_6_3_2","DOI":"10.1021\/ja00142a004"},{"doi-asserted-by":"publisher","key":"e_1_2_6_4_2","DOI":"10.1021\/cm0202557"},{"doi-asserted-by":"publisher","key":"e_1_2_6_5_2","DOI":"10.1016\/j.jssc.2004.07.046"},{"doi-asserted-by":"publisher","key":"e_1_2_6_6_2","DOI":"10.1016\/S0003-2670(98)00152-4"},{"doi-asserted-by":"publisher","key":"e_1_2_6_7_2","DOI":"10.1366\/000370205774430945"},{"doi-asserted-by":"publisher","key":"e_1_2_6_8_2","DOI":"10.1016\/j.jallcom.2003.11.116"},{"doi-asserted-by":"publisher","key":"e_1_2_6_9_2","DOI":"10.1039\/B418964G"},{"doi-asserted-by":"publisher","key":"e_1_2_6_10_2","DOI":"10.1021\/ja020836x"},{"unstructured":"\u00a0","key":"e_1_2_6_11_2"},{"doi-asserted-by":"publisher","key":"e_1_2_6_12_2","DOI":"10.1002\/ange.200353169"},{"doi-asserted-by":"publisher","key":"e_1_2_6_12_3","DOI":"10.1002\/anie.200353169"},{"unstructured":"\u00a0","key":"e_1_2_6_13_2"},{"doi-asserted-by":"publisher","key":"e_1_2_6_14_2","DOI":"10.1002\/1521-3757(20021202)114:23<4681::AID-ANGE4681>3.0.CO;2-D"},{"doi-asserted-by":"publisher","key":"e_1_2_6_14_3","DOI":"10.1002\/1521-3773(20021202)41:23<4495::AID-ANIE4495>3.0.CO;2-I"},{"doi-asserted-by":"publisher","key":"e_1_2_6_15_2","DOI":"10.1039\/b008293g"},{"key":"e_1_2_6_16_2","doi-asserted-by":"crossref","first-page":"1506","DOI":"10.1093\/clinchem\/37.9.1506","volume":"37","author":"Templeton E. 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