Abstract: A method of determining a concentration of analytes of interest in a sample reaction mixture is disclosed. The method can include measuring intensities of the polarized fluorescence of at least one comparative reaction mixture containing different, known amounts of the analytes of interest, and measuring the intensities of the polarized fluorescence of a sample reaction mixture. The method can also include determining the preliminary concentrations of the analytes of interest in the sample reaction mixture by comparing the measured intensities of the sample reaction mixtures at various time points. The margin of error for the preliminary concentration of the analytes of interest in the sample can be determined at the various time points. Finally, the concentration of the analytes of interest in the sample reaction mixture can be determined by comparing the preliminary concentrations and the respective margin of error at the given time points.

Abstract: A method for reducing non-specific binding in an assay is provided herein. The method includes (a) providing a reaction mixture, which includes or is suspected to include a first component and a second component capable of binding to each other in a specific binding reaction, and (b) adding non-physiological amounts of at least one additive to the reaction mixture before, during or after binding in a sufficient amount to reduce non-specific binding in the reaction mixture. The method further includes (c) monitoring or measuring the presence and/or concentration of at least one of the first and second components after step (b).

Abstract: A method of determining a concentration of analytes of interest in a sample reaction mixture is disclosed. The method can include measuring intensities of the polarized fluorescence of at least one comparative reaction mixture containing different, known amounts of the analytes of interest, and measuring the intensities of the polarized fluorescence of a sample reaction mixture. The method can also include determining the preliminary concentrations of the analytes of interest in the sample reaction mixture by comparing the measured intensities of the sample reaction mixtures at various time points. The margin of error for the preliminary concentration of the analytes of interest in the sample can be determined at the various time points. Finally, the concentration of the analytes of interest in the sample reaction mixture can be determined by comparing the preliminary concentrations and the respective margin of error at the given time points.

Abstract: A method of lowering the rate of a specific binding reaction in an assay for the detection and/or measurement of an analyte of interest is provided herein. In particular, the method includes providing a fluorescent conjugate of the analyte; a component capable of specifically binding to the analyte and its fluorescent conjugate; and a sample, which includes or is suspected to include the analyte. The method also includes allowing the specific binding component to interact simultaneously or at different times with the fluorescent conjugate of the analyte and the analyte in the sample, thereby forming a detectable complex due to the reaction between the fluorescent conjugate of the analyte and its specific binding component, wherein the reaction is performed in the presence of non-physiological amounts of at least one additive. The method further includes monitoring for the rate of change of the concentration of the detectable complex as a function of the amount of analyte in the sample.

Abstract: A method of lowering the rate of a specific binding reaction in an assay for the detection and/or measurement of an analyte of interest is provided herein. In particular, the method includes providing a fluorescent conjugate of the analyte; a component capable of specifically binding to the analyte and its fluorescent conjugate; and a sample, which includes or is suspected to include the analyte. The method also includes allowing the specific binding component to interact simultaneously or at different times with the fluorescent conjugate of the analyte and the analyte in the sample, thereby forming a detectable complex due to the reaction between the fluorescent conjugate of the analyte and its specific binding component, wherein the reaction is performed in the presence of non-physiological amounts of at least one additive. The method further includes monitoring for the rate of change of the concentration of the detectable complex as a function of the amount of analyte in the sample.

Abstract: A method for reducing non-specific binding in an assay is provided herein. The method includes (a) providing a reaction mixture, which includes or is suspected to include a first component and a second component capable of binding to each other in a specific binding reaction, and (b) adding non-physiological amounts of at least one additive to the reaction mixture before, during or after binding in a sufficient amount to reduce non-specific binding in the reaction mixture. The method further includes (c) monitoring or measuring the presence and/or concentration of at least one of the first and second components after step (b).

Abstract: The invention provides methods for detecting an analyte in a sample including the steps of: (a) exciting a sensitizer label on an analyte; (b) permitting energy from the excited sensitizer label to be transferred to and excite an acceptor molecule, whereby the sensitizer label returns to an unexcited state; (c) reacting the excited acceptor molecule with a chemiluminescent precursor to form a chemiluminescent compound which emits light in response to an activation source; (d) exposing the chemiluminescent compound to the activating source to produce a detectable signal; (e) detecting the signal; and (f) correlating the signal with the presence or absence of the analyte. The chemiluminescent precursor is desirably an olefin capable of being converted to a 1,2-dioxetane. Target amplification techniques, such as PCR, may be used to directly label a target analyte with a sensitizer.

Abstract: The invention provides chemiluminescent assays that incorporate a film including at least one chemiluminescent precursor immobilized therewith which produces a triggerable chemiluminescent compound, the film being free of compounds which generate singlet oxygen and being adapted for use with a sensitizer-labeled agent or agent probative of the analyte.

Abstract: The present invention relates to chemiluminescent assays which incorporate a second film or membrane which includes a solid chemical component for activation of a stable dioxetane. Decomposition of the stable dioxetane can be accomplished using a combination of heat and chemical treatment.