A) A cation exchange resin. Please explain your answer (3 pts). B) An anion exch

Question

A) A cation exchange resin. Please explain your answer (3 pts).

B) An anion exchange resin. Please explain your answer (3 pts).

C. You separate the same mixture using 2D gel electrophoresis. The isoelectric focusing step of the separation uses a decreasing gradient of pH from 11 to 2. Please draw the final result of the 1st and 2nd steps of the 2D gel electrophoresis in the box below. Please indicate the orientation of the gel in terms of decreasing molecular weight and decreasing pI. Please label the relative location of each protein migration. (3 pts)

Explanation / Answer

We start the chromatography at pH 11. At this paticular pH, all the protine in the mixture are negatively charged.

Cation exchange resin- positively charged proteins (cations) are attracted to the negatively charged resine.In such cases, proteins may adsorb to cation exchangers at pH up to pI+1..the oder is

Anion exchange resin- negatively charged proteins (anions) are attracted to the positively charged resine.In such cases, proteins may adsorb to cation exchangers at pH up to pI-1..

All the protine will bind the resin. As we decrease the pH, the protine will progressively elute when the pH of the buffer becomes lower that their pI.

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