Abstract: A proximity binding assay (PBA) is performed on at least one test sample, at least one reference sample, a background sample, and one or more calibration samples using a thermal cycler instrument. Ct values are determined for at least one set of test sample data and at least one set of reference sample data. Background corrected Ct values are calculated using a corresponding value in a background sample data set. A linear range is determined for the background corrected Ct values as a function of sample quantity. A linear regression line is calculated for each linear range. One or more parameter values of an exponential model (EM) fold change formula are estimated from the one or more sets of calibration sample data. A target protein quantity and associated confidence interval are calculated using the linear regression lines and the EM fold change formula.

Abstract: A proximity binding assay (PBA) is performed on at least one test sample, at least one reference sample, a background sample, and one or more calibration samples using a thermal cycler instrument. Ct values are determined for at least one set of test sample data and at least one set of reference sample data. Background corrected Ct values are calculated using a corresponding value in a background sample data set. A linear range is determined for the background corrected Ct values as a function of sample quantity. A linear regression line is calculated for each linear range. One or more parameter values of an exponential model (EM) fold change formula are estimated from the one or more sets of calibration sample data. A target protein quantity and associated confidence interval are calculated using the linear regression lines and the EM fold change formula.

Abstract: The present invention provides novel, water-soluble, red-emitting fluorescent rhodamine dyes and red-emitting fluorescent energy-transfer dye pairs, as well as labeled conjugates comprising the same and methods for their use. The dyes, energy-transfer dye pairs and labeled conjugates are useful in a variety of aqueous-based applications, particularly in assays involving staining of cells, protein binding, and/or analysis of nucleic acids, such as hybridization assays and nucleic acid sequencing.