As for these samples, the concentration obtained from the standard curve must be multiplied by the dilution factor when analyzing the results. Step 4: Once you click Add-Ins at the bottom, you will see the Manage drop-down list. If your Excel is not showing this pack, follow the below steps to unleash this option. If the OD value of the sample is out of upper limit of the standard curve in sandwich(or out of the lower limit of competitive ELISA), these samples should be diluted before proceeding with the ELISA to obtain an accurate result. Unleash Data Analysis Tool Pack in Excel. Select the 4th work sheet (Fit NLF Find X from Y1), input or copy the OD value of the sample to the left column, the concentration will be calculated automatically. and the pH needs to be set before ELISA analysis considering the high water content and the acidic pH of these materials. Step 6: Calculate the concentration according to the curve Calculated ELISA data for the limit of the detection and quantification with. Then double click the X axis and Y axis to set them to log type. Step4: Check the information of the standard curve (parameter values, R-Square, etc.)ĭouble click the Fitted Curve Plot in step 4 to make the curve to editable state. Select the value box, and then select items in turn as below.Īfter setting the above items, click the Fit button, then the curve and its information is produced as showed in step 4. Step 1: Input information of the OD value and concentration of the standards The software Origin is recommended to use. Plot a four parameter logistic curve on log-log graph paper, with standard concentration on the x-axis and OD values on the y-axis. Subtract the average value of zero standard OD (this step is unnecessary in procedure of competitive ELISA). ELISA samples are always tested in duplicates or in triplicates, then average the absorbance values for each setĢ. As the last procedure of ELISA, calculation of results is of great importance to the success of the assay.ġ.
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