ELISA Analyzer
This software will allow you to conduct 4 parameter logistic (4PL) regression on sigmoid standard curve data and transform sample data to concentrations. This is in particular useful for ELISA data, but other data based on sigmoid-type standard curves are equally applicable.
The software will provide the 4 parameters for the 4PL-regression; Amin, Amax, Hill curve and EC50, which provides the least-squares best fit model of the standard curve data based on the following equation.
Isolating the Concentration from this equation leads to the following formula
The latter equation allows to calculate the concentration in a sample based on the absorption measured. The web application below will automate this process on your data.
You can download a set of sample data that will allow you to test the application here. In the excel file you will find a guide for the application (the guide is also available below the application).
Application author: Martin LARSEN
Guide:
Create excel sheet with the raw data to be analyzed (background subtracted if required). The data should be in a 8x12 matrix with an additional row and column as margins including row and column labels
More than one plate can be included on the same excel sheet with a separator specifying the plateID. The plateID should be placed in the cell on the top-left corner of the matrix (cf. sheet "IgA") and each plate should be separate with an empty line
Layout data
Create excel sheet with the plate layout. The sheet name should be identical to sample data but extended with "_Layout".
The layout should be identical to the sample data format.
Standard curve values should be marked with yellow background color and labelled with a name (e.g. antigen name without spaces) and separate from the concentration with a space (e.g. "IgA 250"). No units should be used
Sample values should be marked with a sampleID (no spaces) and separated from the dilution factor with a space (e.g. "Sample1 500" = Sample1 diluted 500 times)
Standard curve
The standard curve allows you to select range of OD values that you consider within the analytical range. Sample OD values within the range will be translated to the corresponding concentration.
Output
Each sample is displayed with the raw OD value and the corresponding concentration based on the standard curve and the dilution factor.
The concentration is transformed from ng/ml to mg/ml (so divided by 10^6). This part needs to be updated to accommodate any type of ELISA data.
Web server administrator: Paul CASSAN