This strongly depends on the expected volumes, tissue material (amount/weight) etc. I would propose to consult adequate literature sources (e.g. "experimentator" series (Springer) etc.)

Alternative sources:

"Wilson and Walker's Principles and Techniques of Biochemistry and Molecular Biology" (Cambridge University Press; 2018)

or

"Basic methods in molecular biology" (Elsevier, 1986)

etc.

Hi Joao,

Can you clarify your question more?

In general, WB depends on many factors including but not limited to

- how much protein has been loaded

- first and secondary antibody types, specificity, and concentration

- blocking buffer type (milk or BSA ), concentration (3% or 5%), and the time of incubation (overnight or few hours)

- washing buffer

- development reagents and methods