From an SDS-PAGE gel, how can you tell that you successfully induced GFP expression?
Is that related to amount of bands?
If you have made fusion protein of suppose protein 'A' with GFP, then definitely your protein 'A-GFP will be now bigger in size than original protein 'A'' and its molecular weight will be higher. Now If you are using SDS-PAGE, and you are loading protein 'A without GFP' in one well of gel and Protein A-GFP in another well, the well with A-GFP will show band upper to the band corresponding to protein 'A' in another well. This is because when a protein is large sized, it migrates slowly to the bottom than the protein which is bigger in size. Here amounts of bands will not change as long as if the question is related to fusion protein
From an SDS-PAGE gel, how can you tell that you successfully induced GFP expression? Is that...
what would happen to the proteins in your gel if you did not add SDS-PAGE?
1. Figure I shows an SDS-PAGE gel. A) Rank the 3 proteins by size, from largest to smallest. Explain why this trend is observed in SDS-PAGE gels. B) What is the purpose of SDS in SDS-PAGE? C) Sample L is the ladder. What is its purpose? D) Typically, PA (polyacrylamide) is used as the gel for protein electrophoresis, whereas agarose is used for DNA electrophoresis. Explain why a different gel material is used, Specifically referring to the pore size of...
parts a,b, c please
3. Sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis a. After pouring the SDS polyacrylamide gel, you realize that instead of 2 ml, you used 4 ml of 30% acrylamide/bisacrylamide solution for the preparation of the separating gel (in both cases for 5 ml, total gel volume). How would this affect the separation of your proteins during SDS PAGE? Explain your answer! b. You have to remake the gel and are now making sure just to add...
You start running an SDS-PAGE gel at 200V. After 10 minutes, you notice that the dye front has not moved very far and the current has dropped to almost zero. What's the most likely explanation for what has happened here?
SDS Page Gel:
The provided standard protein sample for electrophoresis
consists of 9 polypeptides with molecular weights ranging from 250
to 15 KDa.
Sample 1: Protein A in a sample buffer with
B-Mercaptoethanol
Sample 2: Protein A in a sample buffer without
B-Mercaptoethanol
Sample 3: Protein B in a sample buffer with
B-Mercaptoethanol
Sample 4: Protein C in a sample buffer without
B-Mercaptoethanol
Use the picture below & the information about the proteins
above to answer the following questions.
1a....
1d. Finally, you run SDS-PAGE in the presence of 2-mercaptoet urun SDS-PAGE in the presence of 2.mernantoethanol on a subsample of each peak from each column in parts 1a and ic. The image below shows the migration of molecular weight markers and labels for each sample lane Indicate the location of protein band(s) for each of your samples. Identify the protein(s) in each band in each lane and explain your reasoning Anion Exchange 12 Gel Fitration 12 Gel filtation after...
. In preparation of gel electrophoresis, how many grams of SDS do you need to prepare for 2 grams of protein?
1. Although you are making 12% SDS – PAGE gels, other percentages can be made. a. When would you want to use a 10% gel? b. When would you want to use a 20% gel? 2. Why must you use both acrylamide and bis – acrylamide to make your gel? 3. There are three ratios of acrylamide:bis – acrylamide that are used when making these types of gels. a. What is the primary application for a gel made up of...
What type of information can be gained from an SDS-PAGE? Thanks!
Your friend decides to place Green Fluorescent Protein (GFP)
under the control of the promoter from the lacZ gene we discussed
in class. She put this expression plasmid into a bacterium. The
promoter from the lacZ gene is diagrammed to the right.?
2) Your friend decides to place Green Fluorescent Protein (GFP) under the control of the promoter from the lacZ gene we discussed in class. She put this expression plasmid into a bacterium. The promoter from the lacZ gene...