Sample Data comparing standard curve values using 4-PL vs Linear regression. Please keep in mind that values that are represented at the low end of your curve will not be as reliable as values at the higher end of your curve. Note: Linear regression on Excel should only be used if software that can create a 4-Parameter (4-PL) logistic curve fit is unavailable. Where G7 is your OD value (select the appropriate cell for your data) You can do this by using a calculator and manually solving for "X" or by inputting a formula as shown. This document shows how to analyse ELISA data in R. As long as you know how to function out the data, you wont need a 4 parameter regression. Now that you have your equation, you can solve for "x" and enter your sample OD values as Y. If you have to enter the data into an analysis tool, then I would go with Excel. Go to the "Options" tab on the top of the window and Select "Display equation on chart" and "Display R-squared value on chart"Ĭlick "OK" to view your chart with equations. Select Linear under "Trend/Regression type". Click on the "Chart" tab and on the top menu bar click on "Add Trendline".Ī "Add Trendline" window will appear. Highlight data points by clicking on one of them. Click on the "Scale" tab on the top of the window. Place your chart on a new Excel sheet or insert into your current worksheet.ĭouble click on the x-acis values and a "Format Axis" window will pop up. Once you customize the chart to your preferences, click "Next". Name your Series or add another Series if you want to compare Standard CurvesĪdd information to your chart such as titles and axis labels.Make sure that your X Values are your concentrations and your Y values are your Average OD values.Highlight your data range and click on the "Chart Wizard" icon in the excel tool bar.Ĭlick on "Series" located at the top of the Chart Wizard Window. Note: change the cell location from A2 and B2 to the desired cell location of each Standard OD.Įnter the Standard concentrations that each OD should represent next to their corresponding OD average. Take the averages of the duplicate values either by adding them up and dividing by two on a calculator, or entering the formula =AVERAGE(A2 B2). Overall it can assist researchers to speed up data analysis, reduce computational errors, improve the presentation of analysis by creating charts and distinguishing positive samples shown in red color from the negative samples that appear in blue cells of plate, calculate the threshold and ultimately determine the most economic proportions of antibody and conjugate in the mixture considering the antibody and conjugate prices.If software capable of generating a 4-Parameter Logistic curve fit is unavailable, a standard curve using linear regression analysis in excel can be used.Īfter Optical Densities (ODs) have been collected, enter each standard OD into a worksheet as shown. Users can determine better dilution of the antibody and the conjugate that was extracted from blood of animal for other ELISA tests. The software can also help finding appropriate reagent dilutions for the ELISA reaction. ELISA Analyzer was developed with a user friendly interface to streamline titration and analysis of the data obtained from ELISA and improve rapid and lucid presentation of the results. Accurate and precise calculations of the data is time consuming and tedious. ELISA data is usually handled in the MS Excel worksheets through different methods and different formulas.
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