help with this chem question: A and B have the following pI values: Protein A ;p
ID: 784441 • Letter: H
Question
help with this chem question:
A and B have the following pI values:
Protein A ;pI value 6.2
Protein B ; pI value 7.3
You want to use a phosphate buffer at pH 7.0 separate proteins by ion exchange.
1a)Is it possible to separatre A and B under these buffer conditions? Why or why not? What type of ion exchnage resin (cation or anion) would you use?
1b)DRAW a column profile for the separation using the resin you chose.Be sure to indicate where each of the protiens elutes on the profile?
1c)Deduce the 4th structure of A and B based on the SDS-IMAGE gel"
STDS PROTEIN A PROTEIN B
97 ____
68 ____
____
43 ____
____
26 ____
____
14 ____
, assuming the MW values for protein A=160000MW and B=60000 MW.Explain your conculsions based on the details of the SEPARATION method.
Is it possible that either protein has any identical subunits?
If so, how might you determine if that is the case?
Explanation / Answer
Ion exchange chromatography separates compounds according to the nature and degree of their ionic charge.
The column to be used is selected according to its type and strength of charge.
Anion exchange resins have a positive charge and are used to retain and separate negatively charged compounds,
while cation exchange resins have a negative charge and are used to separate positively charged molecules.
since pI1 is more deviatated from buffer solution pH , negative charges will be stronger in this case so, we will use cation exchage resin
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