In my study utilizing anti-Flag antibody to detect a Flag-tagged protein in HepG2 cell line via Western blotting, I have observed multiple bands beyond the expected molecular weight. I seek guidance regarding the potential origin of these additional bands. Could they represent dimerization/oligomerization events manifesting as higher bands, or degradation products yielding lower bands?
How can I substantiate these observations, particularly considering the presumed specificity conferred by the Flag tag?
Lower molecular weight bands could be proteolytic fragments, of course. Higher molecular weight bands could be due to varying degrees of glycosylation. As a test, can you use one or more antibodies that detect the protein of interest rather than the tag?
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