Affinity chromatography protein Purification
Guidelines for the Affinity Purification Lab Report
1) Estimate the molecular weights of the three prominent bands
you see in your thrombin digest lane.
2) You have been provided with two results of a MALDI-TOF analysis.
a) Calculate the actual molecular weight of the three bands by doing a ratio
calculation, based on the observed MALDI MW of the apomyoglobin
standard and the actual MW of the apomyoglobin standard. Ratio =
observed value/actual value.
b) What do you think the 33941.3 peak on p. 102 corresponds to?
c) Compare the molecular weights obtained for your three prominent bands
by using Rf values to the MW obtained by MALDI-TOF. How much discrepancy did you see between the two methods? If you wanted to be as accurate as possible in obtaining the MW of a protein or peptide,
which method would you use and why?
3) In developing this lab, we had to make a decision on what protease to use to
