Mr (KD) 250 30 150 90 ist the proteins in the order of first to the last that el

Question

Mr (KD) 250 30 150 90 ist 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 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. d. 7 891O PH 3 4 5 6 A+B C+D

Explanation / Answer

a. small sized molecules have large retention time thus the order will be: A(largest)-C-D-B(smallest)

b. below its pI a protein (more acidic) has net positive charge and thus bind to negatively charged resin whereas, negatively charged molecule will flow throgh the column thus, the order will be- B-C-D-A

c.

D. First the molecules will be separated on the basis of pI then on the basis of their molecular mass, thus

A-D-C-B pattern will be obtained on basis of pI (horizontally) vertically from top to bottom on basis of size pattern obtained will be- A-C-D-B

Mr A B C D 250 A protein (250 KD) 200 150 C (150 KD) 100 D (50-100 KD AT 90 KD) 50 B (0-50) at 30 0

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