*Parts of a new datasheet*

In order to preserve the integrity of the data and make sure everything is where EasyBound expects it to be, the datasheets are protected by default. However, there is no password so you can unprotect it if you need to. Using any of EasyBound’s function will reprotect the sheet.

**Model selection**

The model selected from the combobox on the commandbar appears at he top of the sheet. A different model can be used on each sheet. The first option on the menu is called Not to be calculated. This option is useful for maintaining copies of the original data with all values.

**Header**

In the header part it’s possible to enter information about the data. Labels for date, labeled ligand, specific activity of the labeled ligand, unlabeled ligand, treatment, and mg protein/tube. It is optional to provide this information. Values for specific activity and mg protein/tube are, however, required for some manipulations of the data.

**Data area**

The data area have columns for entering X-values, total bound values and unspecific bound values. Total bound values can be given in triplicate and unspecific bound values in duplicate.

*Parts of a calculated datasheet*

After the calculation the model used will be shown in the header area, since the drop down menu can be changed without recalculation.

At the end of the calculation a Runs test is performed to see if the data points are randomly distributed around the fitted curve. If this is not the case “Non-random distribution” will be shown.

**Results area**

The result area is inserted between the header and the data area after the data has been calculated.

The first column show the name of the parameters in the selected model. The second column show the calculated parameter values. The next columns show the standard errors of the parameters.

The following column displays a linear Hill coefficient (nH) for some models, the (weighted) sum of squares (SS) and the degrees of freedom (df). Degrees of freedom is equal to the number of datapoints minus the number of parameters.

The last column display the initial estimate for each parameter. The letter in parentheses indicates how the estimates were generated; A automatic, M manual and F fixed.

On the bottom row of the results area is shown the number of iterations needed to find an optimal fit, the number of data points and if weighting was used. The maximum number of iterations is 20, otherwise “Optimum not found” will be shown. If the calculations were interrupted by the user or by an error, this will also be shown on this row.

The result of a model comparison will be shown in the result area.

**Data area**

The title of the left column will be changed according to the model used. The other columns are average total bound, average unspecific bound, specific bound, estimated bound values and standard residuals of the calculated values.

The standard residuals are used to check for outliers and if the values are randomly distributed around the fitted curve.

**Graphs**

The left graph will show the original observed values as dots and the calculated values as a line. The axes titles are determined by the model used.

With most models it is possible to present a linear transformation of the data in an additional graph.