Question

Think about the two methods (the western and dot blot) how they differ, what unique benefits might each technique present? When might one be more useful than the other?

Answer 1

Answer:

By principle, both Western blot and dot blot are use to identify the presence of protein of interest in a complex mixture of proteins using antibodies.

Difference between western and dot blot:

Western blot is time consuming while dot blot is a time saver.

Western blot requires separation of proteins in gel folowed by blotting while in case of dot blot only blotting process is involved.

Western blot is more informative(presence or absence and specific or non-specific) while dot blot helps only in mere detection of protein.(only presence or absence).

Unique benefits:

In western blot, size of the protein can be detected due to the gel separation. Hence it gives information about specific and non-specific binding.

Dot blot helps in rapid detection of a protein of interest in sample.

When to use what:

Western blot can be used if the protein has to be detected from a complex mixture and also to see if there is a non-specific binding of the antibodies.

Dot blot is a much useful technique for detection of purified proteins. Dot blot can be used detection from complex mixtures only if the specificity of the antibodies used are well known.