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Question: suppose you have a mixture of the following four proteins...

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Suppose you have a mixture of the following four proteins that you would like to separate. Protein A is 90kDa in size and has a pl of 5.6 Protein B is 56kDa in size and has a pl of 9 Protein C is 54kDa in size and has a pl of 8.2 Protein D is 60kDa in size and has a pl of 7.5 You suspect C is your target protein and you know it uses ATP as a substrate. Blue sepharose is an affinity based resin that binds proteins that recognize adenine nucleotides such as ATP and NAD 1. Draw a cartoon of an SDS-PAGE gel with these 4 proteins on it. Indicate the position of the Mvw markers 31kDa, 45kDa, 66kDa and 97.4kDa 2. Draw a graph of fraction number (time) vs protein level (Abs aeo) for the separation of the four proteins above using three types of chromatography discussed in class (ion-exchange, size exclusion and affinity). Label which protein you might expect to find in each peak. Indicate the pH as appropriate. What assumptions do you make?
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