Is a smeared band on a Western Blot a valid result?

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  • Thread starter Sunwoo Bae
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Sunwoo Bae
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Homework Statement
I am trying to analyze a western blot result of expression of Keratin Sulfate in variety of cells.
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From this result, the conclusion is that MDCK-siNGT and MDCK-siB4Gal4 resulted in less keratin sulfate, while MDCK-RCA did not produce keratin sulfate at all. Looking at the gel, the band look very smeared, as if nothing was purified prior to blotting. I have noticed a similar looking gel in other articles as well, but is this a valid result?
 

1. Why does a smeared band appear on a Western Blot?

A smeared band on a Western Blot can appear due to a variety of reasons, such as overloading the gel, uneven transfer of proteins, or degradation of the protein sample.

2. Is a smeared band on a Western Blot considered a valid result?

Yes, a smeared band on a Western Blot can still be considered a valid result as long as the experimental controls are run properly and the band can still be interpreted for the presence of the target protein.

3. How can I prevent smeared bands on a Western Blot?

To prevent smeared bands on a Western Blot, it is important to optimize the protein loading amount, ensure even transfer of proteins from the gel to the membrane, and handle the protein samples carefully to prevent degradation.

4. Can a smeared band affect the accuracy of the Western Blot results?

While a smeared band may affect the visual interpretation of the Western Blot, it may not necessarily affect the accuracy of the results if proper controls are in place and the band can still be analyzed for the presence of the target protein.

5. Should I repeat the Western Blot if I see a smeared band?

It may be necessary to repeat the Western Blot if a smeared band is observed, especially if the band cannot be clearly interpreted for the presence of the target protein. However, it is important to troubleshoot the possible causes of the smearing before repeating the experiment.

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