Question

6. Suppose you have a mixture of five proteins listed in the table below: Protein pl Mol. Welght, kDa Homer 4.6 69 Liza 10.7 13 Marge Maggie Bart 5 40 6.4 8.5 7 27 Indicate the order in which the proteins elute from a Bio-Gel p-10 gel-filtration column (starting with the on that elutes first). Is this the best resin to separate all the components of this mixture? Why? If not, what other Bio-Gel resin can you use to separate all the components of the mixture?(10 pts) This requires you look up some information about the size exclusion limits for each type of resin a. You load this mixture on a cation exchange column. The buffer you use for this column is acetate buffer, pH 4.76. List the protein or proteins that will appear in the flow through (will not bind to the column) (5 points) In order to elute those proteins that are immobilized on the column, you then apply a linear salt gradient, with NaCl concentration gradually increasing from 0 to 1M. Indicate the order in which the proteins bound to the column will elute as the salt concentration increases. (5 points) b. i. ii. Repeat the same procedure as in b. But now you use anion exchange column, and the buffer is TRIS (pH8). Answer the same questions as seen in i. andii(10 points) C.

need only part a and C. please and thank you

Answer 1

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