1. Determine Vo from the elution volume of blue dextran and Ve of cytochrome, the unknown and DNP aspartate._x000D_
2. On a single excel graph, plot the absorbance (y-axis) of each fraction as a function of the volume (x-axis) or fraction number keeping in mind the first fractions collected as a single volume. Indicate the wavelength at which each peak was measured. Determine the midpoint of each peak, either by inspection or by the equation for estimation of the arithmetic mean. The data of Blue Dextran, Cytochrome C, and DNP-Aspartate should be in this graph._x000D_
3. Repeat this procedure for the 1 mL of the unknown solution. Calculate the KD for cytochrome c from your data._x000D_
4. Estimation of the molecular weight of cytochrome c and unknown._x000D_
5. The flowing data have been collected from the literature. Using this information prepare a graph of KD (ordinate) as a function of log10 molecular weight for the known proteins. Estimate the molecular weight of the cytochrome c and your unknown in the preparations you examine by interpolation of your estimated KD value on this semi-log plot. Make sure your report includes the KD and MW for cytochrome c and the unknown._x000D_
6. Calculate the value of K in the experiment._x000D_
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