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For the most accurate results we recommend that 4 parameter logistic (4-PL) curve fitting software be used. The object of 4-PL is to find the line equation that best fits the data within a certain set of limitations. Even in immunometric assays, the dose response curve is very rarely a straight line. By choosing a better curve fit for your data, you will ultimately have more accurate returned sample values. If this software is unavailable the user will have to plot their data and calculate by hand. Most microplate readers come with software.