In a WT strain, a unique band at 26KDa is detected with the gsp1 antibody, but when the same strain is tagged with GFP and overexpressed with promoter ADH, a band appears at 50KDa (as expected) with a good amount , but another appears at 26KDa (less intense).
Dominique Liger
Hi there, the simplest explanation is that your GFP fusion is partially cleaved into GFP and your untagged protein. A way to check this is to probe your membrane with anti-GFP. If the hypothesis is right you should also get two bands for GFP fusion and GFP alone.
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