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Use the following SDS-PAGE and Western Blot results for this 2 part question:

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Wize University Biology Textbook > Lab Techniques for Biology

Nucleic Acid Detection [Northern and Southern Blots]

3 Activities

Wize University Biology Textbook > Lab Techniques for Biology

Protein Characterization

4 Activities

Use the following SDS-PAGE and Western Blot results for this 2 part question:
Match the results to either technique:
A.
Western Blot
B.
SDS-PAGE
A
B
More Nucleic Acid Detection [Northern and Southern Blots] Questions:
Practice: Gene Expression
Which technique would allow you to determine whether the transcription from the CFTR gene was compromised in samples obtained from patients with cyctic fibrosis?
Which technique would allow you to determine whether the transcription from the CFTR gene, was compromised in samples obtained from patients with cyctic fibrosis?
Interpreting and drawing Northern blots
Using the following Northern blot, what can we conclude about the gene expression of Hsp1 in high temperatures? Fill in what you predict to be the banding pattern of Hsp1 and 5S RNA levels at 20oC.
You want to measure the rate of Hsp1 mRNA decay at 55oC. Design an experiment to answer that question using a Northern blot as your read-out.
Practice: Gene Expression
Which technique would allow you to determine whether the transcription from the CFTR gene was compromised in samples obtained from patients with cyctic fibrosis?
Which of the following can be resolved using Western blotting?
1. Protein-protein interaction
2. Expression of a protein when induced with a drug
You are running an experiment where you want to determine if in a eukaryotic cell the rate of an mRNA transcript being degraded. Which of the following describes the experiment and technique you will use?
Unicellular organisms have the ability to change their membrane fluidity depending on temperature. A bacteria is able to survive at -10oC. At this temperature, what would you expect to see on a Northern blot probing for desaturase?
A heat shock protein increases in abundance after exposure to 40°C compared to 24°C. You can visually see the increase in band intensity on a Western blot. You treat your cells with MG132 to block the activity of the proteasome as well as treating the cells with antinomycin to block transcription. Looking at your Western blot, the protein abundance of the heat shock protein decreases over time at the same temperature. What is the most likely reason for this observed decrease?
Interpreting and drawing Western blots
Using the following Western blot, what can we conclude about the protein abundance of HSP1?
What would you add to the preceding experiment to measure the rate of HSP1 degradation?
What would you add to the preceding experiment to measure the rate of HSP1 translation?
Interpreting and drawing Northern blots
Using the following Northern blot, what can we conclude about the gene expression of Hsp1 in high temperatures? Fill in what you predict to be the banding pattern of Hsp1 and 5S RNA levels at 20oC.
You want to measure the rate of Hsp1 mRNA decay at 55oC. Design an experiment to answer that question using a Northern blot as your read-out.
You are studying an olfactory receptor in two strains the roundworm C. elegans. In the wildtype strain, the worms are able to smell attractive food odors. In the mutant strain, the worms are unable to sense when food is nearby. You believe that a severe DNA mutation has led to a deletion of the olfactory receptor gene from the mutant C. elegans worm's genome. How would you detect whether this specific DNA sequence is present in the mutant C. elegans genome?
Which technique would allow you to determine whether the transcription from the CFTR gene, was compromised in samples obtained from patients with cyctic fibrosis?
Which technique would allow you to determine whether the transcription from the CFTR gene, was compromised in samples obtained from patients with cyctic fibrosis?
You are studying an olfactory receptor in two strains the roundworm C. elegans. In the wildtype strain, the worms are able to smell attractive food odors. In the mutant strain, the worms are unable to sense when food is nearby. You believe that a severe DNA mutation has led to a deletion of the olfactory receptor gene from the mutant C. elegans worm's genome. How would you detect whether this specific DNA sequence is present in the mutant C. elegans genome?
In Northern blotting, electrophoresis is used to resolve which biological molecules? What type of probe is used to identify the target molecule(s)?
Source of this question: Griffiths AJ, et al. An introduction to genetic analysis. 11th Ed.
More Protein Characterization Questions:
Practice: SDS-PAGE
Use the following SDS-PAGE results for this 4 part question:
Practice: Detecting Proteins
You are working in a research lab studying a novel protein called WIZE. You have two versions of the protein: the standard version (30 kDa) and a version that has been post-translationally modified with a single ubiquitin molecule (which adds ~8.5 kDa to its mass). You accidentally forgot to label your tubes. What is the best and most direct technique to determine which tube contains the larger, ubiquitinated version of WIZE?
Practice: SDS-PAGE
Use the following SDS-PAGE results for this 4 part question:
Use the following SDS-PAGE results for this 4 part question:
Organize the steps that take place for SDS-PAGE leading into Western Blotting:
The HES1 protein is a bHLH protein that is known to bind to certain gene regulatory motifs and repress transcription. You notice that when HES1 is mutated, the protein Ngn3 is up-regulated. How would you confirm that Hes1 associates with the Ngn3 transcript?
Which of the following protein purification methods can be used to isolate the phosphorylated form of a protein?
How is chromatin immunoprecipitation (ChIP) different from regular immunoprecipitation (IP)?
You are studying a mutated form of Protein X, which has one missense mutation in it's DNA sequence. This mutation results in an amino acid substitution from an Ser to a Tyr. The mutated Protein X is folded into the same native conformation as the wildtype protein and has the same biological function. Which of the following is the best way to separate these two proteins?
Which of the following methods requires the use of antibodies generated against the protein of interest?
Practice: Detecting Proteins
You are working in a research lab studying a novel protein called WIZE. You have two versions of the protein, WIZE-14, which has incorporated light nitrogen, N14 and WIZE-15, which has incorporated heavy nitrogen, N15. Otherwise, they are exactly the same with the same folding structure and biological activity. One morning, you come into the lab and realize that you accidentally forgot to label your tubes last night. Whoops! What is the best technique to determine which tube has WIZE-14 and which tube has WIZE-15?
You learned that the NLS protein tag is both necessary and sufficient for nuclear localization of a protein. Let’s apply this concept to proteins that localize to the ER lumen. When answering the following questions, pick from the following:
Start transfer sequence
ER retention signal
Membrane Proteins
Explain how the following experiments can be used to determine the domain structure (or “topology”) of a membrane-associated protein in conjunction with SDS-PAGE.
a. Treatment with a protease.
b. Treatment with a detergent.