Exploring Mutations in Enolase Gene Expression: A Western Blot Analysis

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In summary, the conversation discusses the presence of two different markers for proteins A and B in the western blot of a patient. This can be explained by changes in genes, such as a nonsense-mutation resulting from a substitution or insertion/deletion of base-pairs. It is also possible that there is a co-dominant expression of two different proteins. In this case, the markers are related to the protein Enolase, with marker B being the normal version and marker A being the mutated version. Other accurate possibilities for mutations and their causes related to this enzyme are also mentioned.
  • #1
hellllo
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Hello Forum,

please have a look at this image

http://imageshack.us/photo/my-images/98/0bigg.jpg/

There are two proteins A and B in the right column.
Marker B is the protein´s length of healthy people.
But as you can see there are two different markers for this protein (the western Blot was made for one patient).

How can you explain the appearance of two different markers in one patient?

Can you explain with examples how changes of genes can lead to this result?
 
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  • #2
This sounds like a homework problem. I can think of several explanations, what do you think could have happened?
 
  • #3
Can you explain with examples how changes of genes can lead to this result?

The answer to this question is a nonsense-mutation because of the substitution of one base-pair (exmaple 1). The second example is a nonsense-mutation because of insertion or deletion of base-pairs.

Do you have any further examples which explains this result?

How can you explain the appearance of two different markers in one patient?

The different genes express two different proteins. The "healthy" gene codes for the "healthy" protein and the other gene codes for the short protein. Both proteins are expressed in this patient.
Mutation is caused due to radioactive waves or it is inherited. It is possible that there is a co-dominant expression and, therefore, both proteins are expressed.

Do you have any other possibilities?

The markers above are related to the protein Enolase. Marker B is the Enolase and marker A is the mutated enzyme.

Maybe you have some more accurate possibilities considering the mutations and their causes which are related to this enzyme.
 

1. What is a Western blot?

A Western blot is a laboratory technique used to separate and detect specific proteins from a complex mixture. It involves separating proteins by size using gel electrophoresis and then transferring them onto a membrane for detection.

2. What is the purpose of a Western blot?

The purpose of a Western blot is to identify and quantify the presence of specific proteins in a sample. This can be used to study protein expression levels, protein-protein interactions, and post-translational modifications.

3. How does a Western blot detect two proteins?

In a Western blot, the proteins are separated by size and then transferred onto a membrane. The membrane is then probed with specific antibodies that bind to the target proteins. The bound antibodies can then be visualized using a detection method, such as chemiluminescence, to identify and quantify the proteins of interest.

4. What is the difference between a one-protein and two-protein Western blot?

A one-protein Western blot involves detecting and quantifying a single protein of interest, while a two-protein Western blot involves detecting and quantifying two different proteins simultaneously. This can be done by using two different primary antibodies, each specific for one of the target proteins, and using secondary antibodies labeled with different fluorophores or enzymes for detection.

5. What are some potential challenges with performing a Western blot for two proteins?

Some potential challenges with a two-protein Western blot include optimizing the antibody concentrations and detection method to ensure equal detection of both proteins, as well as avoiding cross-reactivity between the two proteins. Additionally, the efficiency of the protein transfer and potential sample degradation during the process can also affect the accuracy of the results.

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