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2nd dim: high-weight zone depleted

Hi everybody,
I encounter a big main problem in 2-dim electrophoresis: all the high weight zone is depleted, like "erased" from the gel. It happens indipendently from the protein quantity and actually I suspect it is a problem of the 2nd dimension: of 12 tubes I ran for the 1st dimension, the first 4 SDS-PAGE were fine, the other 8, plus others I ran afterwards, present the same problem. I try to upload a phote, maybe it helps. Any hint will be welcome, I am currently stacking on this!
:worshippy:

Re: 2nd dim: high-weight zone depleted