During the purification of a mixture of proteins listed in the table below: Name
ID: 53355 • Letter: D
Question
During the purification of a mixture of proteins listed in the table below:
Name MW (kDa) pI
A 28 5.5
B 19 6.0
C 24 8.0
D 29 5.2
E 58 9.5
F 35 4.3
G 200 6.5
H 94 7.5
I 62 6.0
a. Which proteins bind to a CM-cellulose column running at pH 7.0?
b.Which protein will run through a Superdex200 gel filtration column first? Which one will come out last?
c. Which protein will be eluted last from a DEAE-cellulose column at pH 7.0?
d. Protein G runs on a SDS-PAGE gel as a 50 kD band. How many polypeptide chains make up this protein? Estimate how many residues this polypeptide chain contains.
e. Propose an effective purification strategy for protein E.
Explanation / Answer
a. C 24MW pH -8.0, E 58MW pH 9.5, H 94MW pH 7.5
b. G 200MW will run first and B 19MW will run last
c. G 200MW pH 6.5
d. three domain with 3 polypeptide chains, 2 residues
e centromere protein E, 312kDa can be purified by affinity purification
Affinity Purification (AP), Antibody Array (AA), ELISA, Western Blotting (WB)
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