Homework 9- Due 11-13 (2)
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Dec 6, 2023
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Homework 9
Due Nov 13
th
10 points
Homework 9
Activity 1
: Bradford Assay
Please read a short information sheet from Bio-Rad, and answer the following questions.
https://www.bio-rad.com/featured/en/bradford-assay.html
1.1
There are two commonly used Coomassie dyes: Coomassie G-250 and Coomassie R-250
dyes. Which one is used in Bradford Assay and which one is preferred for staining protein
gels?
(2 points)
1.2
Coomassie G-250 can exist in three forms: cationic (protonated), neutral and anionic
(unprotonated). Which form of the dye is present in the Bradford Protein Assay Reagent?
Which form is the dye in when bound to the protein?
(2 points)
1.3
What problem are you likely to run into if the concentration of your Unknown protein is
higher than the concentration of the standard protein dilutions used for building the
standard curve? What solution to this problem can you offer?
(2 points)
Activity 2
: SDS-PAGE Animation Video. Please watch the video and answer the following
questions.
https://www.youtube.com/watch?v=i_6y6Z5UvwE
2.1
What is the purpose of SDS in protein sample buffer and SDS-PAGE?
(1 point)
2.2
What is the role of bis-acrylamide in polyacrylamide gel?
(1 point)
2.3
What is the purpose of the stacking gel in SDS-PAGE?
(1 point)
2.4
What is the purpose of APS and TEMED in polyacrylamide gel casting?
(1 point)
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