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. 2012 Mar 28;95(5):1096–1102. doi: 10.3945/ajcn.111.032862

FIGURE 3.

FIGURE 3.

Identification of the protein nonspecifically detected by the human ZIP8 antibody. A: Detection of IgG incorporated into the protein sample during immunoprecipitation with the ZIP8 antibody (α-ZIP8) by an anti-IgG secondary antibody (α-IgG). B: Position of the zinc-responsive band nonspecifically produced by α-ZIP8 band on a Western blot. The nonspecific band is indicated by an asterisk. C: Coomassie stain of a counterpart gel of the Western blots shown in panels A and B. The region migrating at the size of the nonspecific protein is indicated by a dashed box and was excised from the stained gel for LC-MS/MS. D: The peptide profile of the isolated proteins determined by LC-MS/MS. Dematin, with an expected molecular weight of 46 kDa, showed the highest normalized spectrum count among the 11 identified proteins. E: Peptide sequences unique for dematin identified by LC-MS/MS are highlighted in gray. HC, heavy chain; IP, immunoprecipitation; LC, light chain; LC-MS/MS, liquid chromatography–tandem mass spectrometry; ZIP8, Zrt/Irt-like protein 8.