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suppmat_1583_supp1.ppt933KBiotinylation of Cytochrome C was assessed following the binding of HRP-conjugated streptavidin and detection by ECL (8% SDS-PAGE). Lane 1 shows no reactivity with native cytochrome c and Lane 2 shows heavy reactivity with biotinylated cytochrome c.
suppmat_1583_supp2a.ppt35KReactivity of positive control sera. Anti-self-PDC serum (diluted 1:1000) was tested in ELISA against native and modified antigen preparations.
suppmat_1583_supp2b.ppt1766KImmunoreactivity of positive control serum (1:1000) with KLH-B (Lane 5), KLH (Lane 6), mP/O-B (Lane 7) and mP/O (Lane 8). Lanes 1-4 represent the protein stained membrane corresponding to contents of Lanes 5-8.
suppmat_1583_supp3.ppt38KAntibody reactivity at 6 weeks (diluted 1:1000) against native and modified Cytochrome C following sensitisation of animals with KLH-B. This shows the reactivity is wholly biotin related.
suppmat_1583_supp4.ppt2110KBiotin pre-absorption. Sera from KLH-B and mP/O-B sensitised animals were diluted 1:1000 and incubated without (-) or with (+) NHS-LC-Biotin. Samples were then probed against native mP/O and bound antibodies detected using anti-mouse IgG-HRP conjugate. Note reactivity can be totally absorbed from the KLH-B sensitised group but not the mP/O-B group. No absorption is seen with positive control antibody.
suppmat_1583_supp5.ppt50Ka) Splenic T-cell proliferative responses to native KLH, biotinylated KLH (KLH-B), native mP/O and biotinylated mP/O (mP/O-B) in mice (n=5) sensitised with KLH. b) Splenic T-cell proliferative responses to native mPDC (mP/O), biotinylated mPDC (mP/O-B), KLH and biotinylated KLH (KLH-B) in mice (n=5) sensitised with mP/O. No significant proliferative response above background (control) was seen for any antigen.
suppmat_1583_supp7.ppt33KAntibody response to KLH (at 1:10000 dilution) elicited by sensitisation by native and biotiny lated KLH.

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