I have a curious result where my protein of interest is significantly down-regulated (90%) in knockout cells by Western blot, but appears to be unchanged on IF. I have repeated the western result many many times, with 12 technical replicates using lysate from 4 different passages.
My plan was to confirm decreased levels by IF and see if there was any change in localization.
The primary is a rabbit polyclonal that is supposed to work for WB, IF, and IHC. Cells were fixed in formalin, permeabilized/blocked, and then stained with 2ug/mL of my primary, followed by 1:500 (goat anti rab -594). For some reason I observe a strong signal in my knockout cells, which I cannot explain.
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