Please make sure you answer it carefully and completely 4. The following questio
ID: 494707 • Letter: P
Question
Please make sure you answer it carefully and completely 4. The following question are based on the following proteins: A TB TC DE 40 80 200 100 6 17 Protein Mr (KD) a. list first to the last that eluted out from a size exclusion column the proteins in the order of b. list the proteins in the order of first to the last that eluted out from a cation exchanger column Predict the relative position of the proteins if they are subjected to SDS-PAGe the in C. following figure d. Draw relative positions of the proteins after the mixture of the four proteins applied to two dimensional gel electrophoresis: isoelectric is in the x is applied in the y-axis. pH 3 4 5 6 7 8 9 10 A+B+C+D Extra credit: Draw the structure of SDS.Explanation / Answer
In size exclusion chromatography, small molecules are diffused throught the pores. large molecules does not fit in any pore and as a result first to elute out.
The order is :
C>D>B>A
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when the pI of a protein is the same as the pH of a solution, the net charge of the protein will be zero. Therefore, when the buffer solution's pH reaches the pI of the protein, the protein's net charge will be zero. The protein will no longer bind to the stationary phase and will be released and washed out.
Order of elution :
B>D>A>C