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. 2012 Nov 28;288(4):2238–2245. doi: 10.1074/jbc.M112.356089

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© 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 7. — Immunological analysis of cruciferin phosphorylation. Protein storage vacuole fractions were resolved by two-dimensional IEF/SDS-PAGE and blotted onto membranes, and blots were developed using antibodies directed against three different phosphorylation sites. As a control, phosphorylated BSA (phosphorylated at tyrosine (■); phosphorylated at threonine (▴); phosphorylated at serine (●)) was separated within the same gel. A, Coomassie-stained reference gel. B, immunoblot developed with an IgG directed against phosphorylated tyrosine; C, immunoblot developed with an IgG directed against phosphorylated threonine; D, immunoblot developed with an IgG directed against phosphorylated serine. Note that the phosphotyrosine-BSA control on C is not indicated because it partially was beyond the limit of the blot. However, the specificity of the IgG was verified by independent experiments (not shown).