How to obtain sharper protein bands in Western blot?

Question: The protein bands on my blots are fuzzy. Is there any way I can improve the blots?

Answer: As proteins are visualized during the last step of the Western blot procedure, how the bands are presented on the membrane, depends on several factors. Please go through the below checklist if you are having issues:

Was the sample been properly solubilized?
Was electrophoresis conducted too fast so that heat was generated?
Was the gel percentage adjusted to the molecular weight (MW) of the protein? I.e. low percentage for proteins of high molecular weight (HMW), and high percentage for proteins of low molecular weight (LMW)?
Was a high-quality protein loading buffer used with a reducing reagent?
Was the sandwich assembled tightly, pressing the gel sufficiently to the membrane? The sandwich should not be too loose.

If you have any further questions about this issue, or would you like to share your experience, contact Agrisera!

Assemnly of a sandwich for wet protein transfer

Recording of Agrisera online Workshop on optimization of Western blot technique.

Agrisera Western blot protocol.

How to obtain sharper protein bands in Western blot?

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