Homework 9- Due 11-13 (2)

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Purdue University *

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Biology

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Dec 6, 2023

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docx

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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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