Figure 3: ● • . ● KDa ● 97.4 66.2 45.0 ● 31.0 21.5 14.4 S-1 p-1 2.8 2-d S.3 p-3 This figure was generated by centrifuging a pura sample of protein, removing the supernatant, and resuspending the pellet in the same volume as the supernatant to allow direct comparison. The supernatant and pellet samples were then prepared for SDS-PAGE identically and run via normal SDS-PAGE procedures. In the figure, "s" means supernatant and "p" means pellet. The cext or number after the dash represents a different condition. For example, s-1 and p-1 are the supernatant and pellet samples under condition 1. It is not shown, but under wild-type conditions, essentially all of the protein is found in the supernatant. P-4 ● What does the intensity of each band represent? ● Would you find soluble protein in the supernatant or pellet? Why? Would you find aggregated protein in the supernatant or pellet? Why? For each condition (there are 5 different conditions), is there a higher percentage of the total protein in the supernatant, pellet, or is it about equal? What does the percentage of the total protein in the pellet tell you about any condition? What is the approximate molecular weight of the protein? ● What does the band at just under 31 KDa represent? Justify your answer. What does the band at just under 45 KDa represent? Justify your answer. What does this assay tell us about aggregation knowing that under wild-type conditions, essentially all of the protein is found in the supernatant? Why is this a useful experiment to perform for a protein especially if you are interested in protein aggregation? s-5 s-d a
Figure 3: ● • . ● KDa ● 97.4 66.2 45.0 ● 31.0 21.5 14.4 S-1 p-1 2.8 2-d S.3 p-3 This figure was generated by centrifuging a pura sample of protein, removing the supernatant, and resuspending the pellet in the same volume as the supernatant to allow direct comparison. The supernatant and pellet samples were then prepared for SDS-PAGE identically and run via normal SDS-PAGE procedures. In the figure, "s" means supernatant and "p" means pellet. The cext or number after the dash represents a different condition. For example, s-1 and p-1 are the supernatant and pellet samples under condition 1. It is not shown, but under wild-type conditions, essentially all of the protein is found in the supernatant. P-4 ● What does the intensity of each band represent? ● Would you find soluble protein in the supernatant or pellet? Why? Would you find aggregated protein in the supernatant or pellet? Why? For each condition (there are 5 different conditions), is there a higher percentage of the total protein in the supernatant, pellet, or is it about equal? What does the percentage of the total protein in the pellet tell you about any condition? What is the approximate molecular weight of the protein? ● What does the band at just under 31 KDa represent? Justify your answer. What does the band at just under 45 KDa represent? Justify your answer. What does this assay tell us about aggregation knowing that under wild-type conditions, essentially all of the protein is found in the supernatant? Why is this a useful experiment to perform for a protein especially if you are interested in protein aggregation? s-5 s-d a
Chapter10: Reconstitution Of Powdered Drugs
Section: Chapter Questions
Problem 7.2P
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Question
Transcribed Image Text:Figure 3:
●
●
●
●
●
●
KDa
●
97.4
66.2
45.0
●
31.0-
21.5
14.4
S-1
p-1
S-2
2-0
This figure was generated by centrifuging a pura sample of protein, removing the supernatant,
and resuspending the pellet in the same volume as the supernatant to allow direct comparison.
The supernatant and pellet samples were then prepared for SDS-PAGE identically and run via
normal SDS-PAGE procedures. In the figure, "s" means supernatant and "p" means pellet. The
text or number after the dash represents a different condition. For example, s-1 and p-1 are the
supernatant and pellet samples under condition 1. It is not shown, but under wild-type
conditions, essentially all of the protein is found in the supernatant.
S-3
● What does the intensity of each band represent?
● Would you find soluble protein in the supernatant or pellet? Why?
Would you find aggregated protein in the supernatant or pellet? Why?
For each condition (there are 5 different conditions), is there a higher percentage of the
total protein in the supernatant, pellet, or is it about equal?
What does the percentage of the total protein in the pellet tell you about any condition?
What is the approximate molecular weight of the protein?
What does the band at just under 31 KDa represent? Justify your answer.
What does the band at just under 45 KDa represent? Justify your answer.
p-3
S-4
t-d
s-5
s-d
a
What does this assay tell us about aggregation knowing that under wild-type conditions,
essentially all of the protein is found in the supernatant?
Why is this a useful experiment to perform for a protein especially if you are interested in
protein aggregation?
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