1. The following question are based on the following proteins: a. list the prote

Question

1. 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 relation position 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

Part-a:

Answer:

Protein D has molecular weight with 200KD first elutes out of column followed by protein C (120KD), followed by protein A (80KD) finally protein B (40KD) elutes out of column

Reason:

Principle of size exclusion chromatography is “adsorption”. Larger molecules with more molecular weight cannot pass through pores within the stationary phase and thereby they elute first followed by smaller molecules with low molecular weight as these molecules are bounded in the porous particles of polymer stationary phase. Therfore, protein D has molecular weight with 200KD first elutes out of column followed by protein C (120KD), followed by protein A (80KD) finally protein B (40KD) elutes out of column

Part-b:

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