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Nancy1 Nancy1
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11 years ago
Can anyone please help me with these lab questions?
These are questions from the lab called: Identification of Serum Glcoprotein by Polyacrylamide Gel Electrophoresis and Albumin Western Blotting.

1. Describe the results of your Coomasie- stained gel picture. What is in each lane?  Do you have multiple bands or single bands? Why?

2. Which lanes had bands in the sample Western Blot that your TA showed you from this experiment? Why?

3. You accidently use an SDS-PAGE system to do this experiment. What impact would this have on your Western blot results?


4. If you wanted to determine the molecular weight of albulmin, how would you change your experimental protocol?

I know it is a lot but my TA is very hard grader if you can help me doing this I appreciate any help. Thanks.
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#1
Valued Member
11 years ago
1) Western blotting identifies with specific antibodies proteins that have been separated from one another according to their size by gel electrophoresis. The Coomasie is used to identify if proteins have migrated uniformly. Notice at the two cm mark, you have a uniform line coming from all wells.

4) You'd use a molecular weight marker. Typically, molecular weight markers are run in the outer lanes of a gel for comparison to test samples that are run in the middle lanes. A standard curve can be constructed from the distances migrated by each marker protein. Then the distance migrated by the sample proteins can be plotted and their molecular weights calculated by interpolation. More often, however, MW markers are used as a reference to help confirm the identity of proteins of interest whose sizes are already known.
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#2
Educator
11 years ago
Hi Nancy, could you give us an update with what you ended up doing?
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