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. Author manuscript; available in PMC: 2017 May 19.
Published in final edited form as: Cell Chem Biol. 2016 May 12;23(5):618–628. doi: 10.1016/j.chembiol.2016.04.006

Figure 3. Peripherin is the antigen recognized by antibodies that bind 4.

Figure 3

A) General experimental flow for antigen discovery using epitope surrogates. B) SDS-PAGE gel following immunoprecipitation of pancreas tissue lysate with serum-derived antibodies that were purified over a sepharose column containing 4. C) Control SDS-PAGE gel after immunoprecipitation of pancreas tissue lysate with control IgG. D) Assessment of serum antibody binding to 4 from healthy and control donors in the absence of competitor (black bars). Competition binding experiments (red bars) were performed using T1D serum in the presence of 500 nM nonspecific competitor (HaloTag protein) or 500 nM peripherin. E) Competition analyses performed on an epitope surrogate 1, that does not compete with 4 for antibody binding. Black bars represent binding in the absence of any competitor protein. Red bars indicate competition of T1D serum using 500 nM of the competitor indicated. In all cases data are reported as the mean ± s.d. from three experiments. Statistical significance was determined using an unpaired t-test: **P < 0.01; ***P < 0.001; ns = not significant.