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Serum samples were collected on day 28 and stored at 35C, as previously described ( 2.5 Hybridoma fusion, screening, and mAbs purification The spleens of selected mice were used to isolate SCMS- or MetE-specific mAbs as previously described ( 2.6 ELISA The antigen specificity of the immunized mice sera and of the hybridoma supernatants was analyzed by ELISA, as previously described ( 2.7 SDS-PAGE and Western blot analysis SDS-PAGE and WB were carried out to analyze recombinant proteins and SCMS, as previously described ( Human sera were analyzed by WB after SDS-PAGE of rMetE: blotted membranes were incubated with sera diluted 1:100 and detected using HRP-conjugated goat anti-human IgG (Sigma-Aldrich) and signals were detected by Crescendo Western HRP chemiluminescent substrate

Generic copper peptide preparations may vary in sequence, copper coordination chemistry, and purity, making them unsuitable as reference compounds in controlled studies