Why bands on the blot are not distinct from each other?

Q: Why can I not see separate bands of a target protein in my samples, but all look like one band on the blot?

A: There can be few reasons, as:

Protein load/well could have been too high, which led to bands merging together, making them difficult to distinguish from each other.
Proteins could form aggregates or complexes, leading to poor band separation.

Remedy: decrease protein load/well and use less sensitive detection reagent.

Why bands on the blot can not be distinguished from each other?

Agrisera detailed Western blot protocol

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Technical blog

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