Quantitative analysis of samples using a Five Parameter Logistic Fit (5PL) suitable for asymmetric sigmoidal data. All samples are first corrected by the mean of the blank group measurements. The standard data points are plotted (concentration vs. corrected measurement) and a Five Parameter Logistic Fit (5PL) is made through these points. The concentrations of the unknown samples are determined from the fit. It is important to note that concentrations can only be determined for samples which fall within the range of the determined upper and lower asymptotes of the fit (the a and d parameters).

Enzyme immunoassay (EIA) for the quantification of insulin in mouse and rat sera. A dose response curve of the corrected absorbance (450nm - 590nm) vs. concentration is generated using a 5-parameter logistic fit. Concentrations of insulin, present in the samples, are determined directly from this curve with any specified dilution factors applied.

Sandwich ELISA for the quantification of Interleukin 10 (IL10). This method plots the absorbance versus cytokine concentration on a log-log chart and performs a 5 parameter logistic curve fit through the standards. The standard curve is used to calculate cytokine concentration of the unknown samples. If samples were diluted the concentration is multiplied by the specified dilution factor. The %CV, Standard Deviation and Standard Error are calculated for each replicated sample. Samples outside the range of the standards or the fit (greater than the upper asymptote or below than the lower asymptote) are highlighted in yellow.

Quantitative analysis of samples using a Five Parameter Logistic (5PL) curve fit suitable for calculating concentrations from asymmetrical sigmoidal calibrators. This analysis optionally includes a background correction step. If a blank group is included on your layout, the mean of the blank replicates is first subtracted from the raw data measurements (the corrected values are then used in the fit). The standard data points (concentration vs. measurement) are plotted on semi-log axes and a 5PL is made through the points. The concentrations of the samples are determined from the fit with any specified dilution factors applied. The %CV, Standard Deviation and Standard Error are calculated for each replicated sample. Samples outside the range of the standards or the fit (greater than the upper asymptote or below than the lower asymptote) are highlighted in yellow.

Quantitative analysis of samples using a Five Parameter Logistic (5PL) curve fit suitable for calculating concentrations from symmetrical sigmoidal calibrators. Data points are weighted using the expresson 1/y meaning that points with a lower signal have a higher weight. This analysis optionally includes a background correction step. If a blank group is included on your layout, the mean of the blank replicates is first subtracted from the raw data measurements (the corrected values are then used in the fit). The standard data points (concentration vs. measurement) are plotted on semi-log axes and a 5PL is made through the points. The concentrations of the samples are determined from the fit with any specified dilution factors applied. The %CV, Standard Deviation and Standard Error are calculated for each replicated sample. Samples outside the range of the standards or the fit (greater than the upper asymptote or below than the lower asymptote) are highlighted in yellow.

Quantitative analysis of samples using a Five Parameter Logistic (5PL) curve fit suitable for calculating concentrations from symmetrical sigmoidal calibrators. Data points are weighted using the expresson 1/y² meaning that points with a lower signal have a higher weight. This analysis optionally includes a background correction step. If a blank group is included on your layout, the mean of the blank replicates is first subtracted from the raw data measurements (the corrected values are then used in the fit). The standard data points (concentration vs. measurement) are plotted on semi-log axes and a 5PL is made through the points. The concentrations of the samples are determined from the fit with any specified dilution factors applied. The %CV, Standard Deviation and Standard Error are calculated for each replicated sample. Samples outside the range of the standards or the fit (greater than the upper asymptote or below than the lower asymptote) are highlighted in yellow.

Quantitative analysis of samples using a Five Parameter Logistic (5PL) curve fit suitable for calculating concentrations from asymmetrical sigmoidal calibrators. The sensitivity or Lowest Detection Limit (LDL) is calculated as 2 SD above the mean of the Zero replicates. This analysis optionally includes a background correction step. If the Blank group is included on your layout, the mean of the Blank replicates is first subtracted from the raw data measurements (the corrected values are then used in the fit). The standard data points (concentration vs. measurement) are plotted on semi-log axes and a 5PL is fitted through the points. The concentrations of the samples are determined from the fit with any specified dilution factors applied. The %CV, Standard Deviation and Standard Error are calculated for each replicated sample. Samples outside the range of the standards or the fit (greater than the upper asymptote or below than the lower asymptote) are highlighted in yellow.

Quantitative analysis of samples using a Five Parameter Logistic (5PL) curve fit suitable for calculating concentrations from asymmetrical sigmoidal calibrators. The sensitivity or Lowest Detection Limit (LDL) is calculated as 3 SD above the mean of the Zero replicates. This analysis optionally includes a background correction step. If the Blank group is included on your layout, the mean of the Blank replicates is first subtracted from the raw data measurements (the corrected values are then used in the fit). The standard data points (concentration vs. measurement) are plotted on semi-log axes and a 5PL is fitted through the points. The concentrations of the samples are determined from the fit with any specified dilution factors applied. The %CV, Standard Deviation and Standard Error are calculated for each replicated sample. Samples outside the range of the standards or the fit (greater than the upper asymptote or below than the lower asymptote) are highlighted in yellow.

Quantitative analysis of samples using a 5PL standard curve fit plotted on log-log axes to calculate concentrations. This protocol is suitable for enzyme-linked immunosorbent assays measured on 96-well (12x8) microplates. This analysis optionally includes a background correction step. If a blank group is included on the selected layout, the mean of the blank replicates is first subtracted from the raw data measurements (the corrected values are then used in the fit). The standard data points (concentration vs. measurement) are plotted and the selected curve is fitted through the standard data points. The concentrations of the samples are determined from the fit with any specified dilution factors applied. Samples outside the range of the standards are highlighted in yellow. The %CV of the final concentration replicate values are computed.