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Immunoblot analysis of GlxA maturation from samples over-expressed in E. coli. GlxA is over-expressed and purified in either Cu-replete conditions (holo-GlxA) or Cu-starved conditions (apo-GlxA). Samples were prepared in SDS–PAGE buffer and heated at 95°C for 3 min. The C121G variant is unable to form the Tyr–Cys cross-link and migrates together with apo-GlxA, which requires the addition of Cu to form the cross-link. Molecular weight markers are indicated in kDa.
