The following question are based on the following proteins: a. list the proteins
ID: 81154 • Letter: T
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The following question are based on the following proteins: a. list the proteins in the order of first to the last that eluted out from a size exclusion column b. list the proteins in the order of first to the last that eluted out from a cation exchanger column c. Predict the relative position of the proteins if they are subjected to SDS-PAGe in the following figure d. Draw relative positions of the proteins after the mixture of the four proteins applied to two dimensional gel electrophoresis: isoelectric focusing is applied in the x axis; SDS-PAGE is applied in the y-axis.Explanation / Answer
a) According to size exclusion chromatography D will be eludted firat followed by C, A and B.Size exclusion chromatography is one of the HPLC separation modes. The column used is filled with material containing many pores. When dissolved molecules of various sizes flow into the column, smaller dissolved molecules flow more slowly through the column because they penetrate deep into the pores, whereas large dissolved molecules flow quickly through the column because they do not enter the pores. Consequently, larger molecules elute from the column sooner and smaller molecules later, which effectively sorts the molecules by size. This is the separation principle of size exclusion chromatography.
c) The sequence of run in the wells will be farthest 'b' then 'a', 'c' and 'd'. In SDS Page, SDS imparts charges to the protein fragments and as the charge of the samples become same they resolute on the basis of molecular weight. The one with lowest Mwt runs faster and with high Mwt runs slowest.
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