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Using regular sterile water (no preservative), shaking instead of swirling, or storing at room temp degrades the peptide fast

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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