Four Parameter Logistic Curve

Quantitative analysis of samples using aFour Parameter Logistic (4PL)curve fit suitable for calculating concentrations from symmetrical sigmoidal calibrators. This method is widely used and cited in data analysis for typical ELISAs.

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 4PL is made through the points. The concentrations of the samples are determined from the fit with any specified dilution factors applied.

Note, the concentration units are unspecified; use whichever units are applicable for your standards (the calculated concentrations will use these same unspecified units).

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 the lower asymptote) are highlighted in yellow.

Four Parameter Logistic Curve (log-log)

You can supply your raw data in two ways:

) the raw data into the edit box. Use the same format as the example data (i.e. numbers separated by spaces).More…

Select a data file on your computer and press

. Most raw data file formats are automatically recognised and processed. Formats not currently supported can be added very quickly.More…

After reviewing other analysis parameters press the

button below to process your results.

If the entire plate was read then supply the measured value for each well. Alternatively, if only specific parts of the plate were read then list only those measured values. In this case an additional step is required: under theMicroplatesection below, select or create a layout which defines which wells were read and which were not.More…

Select option to provide and process measurement data read across multiple plates.More…

To process data from an Excel document, instead of uploading it here, please try one of the following tools:

•MyAssays Excel Add-infor Windows/Apple Mac.

•MyAssays Desktopfor Windows. This tool can automatically import your data from Excel and combine data from multiple Excel files (for multiple plate analysis). Video walk-through ofimporting data from Excel document available here.

option above and then copy and paste your data from Excel into the edit box.

Select how your samples are arranged on the microplate:

Pick a layout corresponding to how your samples are arranged.

6 Standards (with Background Correction)

Review or edit the concentration values here:

the values into the edit box. If the values are in a sequence, select the

The series is calculated from the value specified in the first row and the sequence settings:

To remove the series and enter your own values push the

A zero concentration value has been specified with a logarithmic x axis.

This is valid but means that you will not see the zero standard on the chart, also in certain cases it may result in a skewed fit. One alternative is to use a suitably low value, such as 0.01 instead of zero.More…

Review or edit the dilution factors here:

the values into the edit box. If the values are in a sequence, select the

The series is calculated from the value specified in the first row and the sequence settings:

If the series repeats in blocks, press the restart button

and enter the number of rows in each repeated block.

To remove the series and enter your own values push the

Optionally provide an ID for each of your Unknown samples.

Provide identifiers for each of your samples. If your IDs use a numeric sequence press

The Sample ID series is generated from the specified first ID and increment value.

Optionally provide additional data to store alongside the results for this assay:

Optionally provide a custom run name; this is useful to help you to identify these results in the future. The provided name will appear at the top of the generated report and on your home page (under your analyses list).

Optionally provide notes to store alongside this assay analysis (e.g. Lot ID, preparation notes, etc.). The provided notes will be included at the top of the generated report.

Click here to remove all flags.ResultsProcessing, please wait…

Only members can access this function.

Pleaselog inorjoinnow (its free to join).