| CPC G01N 21/6428 (2013.01) [C07D 311/82 (2013.01); G01N 33/542 (2013.01); G01N 2021/6432 (2013.01); G01N 2021/6441 (2013.01); G01N 2470/12 (2021.08)] | 19 Claims |

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1. A method for determining the presence of or the amount of an analyte in a sample comprising:
(i) providing a sample suspected of containing an analyte;
(ii) contacting the sample with a fluorescent tracer and a binding partner to provide an assay composition;
wherein the binding partner is specific for the analyte and the fluorescent tracer;
(iii) irradiating the assay composition with light at a first wavelength; and
(iv) measuring the intensity of light emitted at a second wavelength,
wherein
(a) the intensity of the light emitted at the second wavelength is greater if the fluorescent tracer is not bound to the binding partner than if the fluorescent tracer is bound to the binding partner,
(b) the light at the first wavelength is not linearly polarized, and
(c) the fluorescent tracer is selected from the group consisting of:
![]() wherein T is a bond or a linking group and E is an epitopic moiety capable of specifically binding to the binding partner; and
wherein the intensity of the light emitted at the second wavelength is directly proportional to the concentration of the analyte in the sample.
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