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.

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.