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- Supplemental Figure 1 - Ab end specific antibody characterization. Synthetic peptides (Ab1-38, Ab1-40 or Ab1-42) were loaded onto ELISA plates overnight, washed, and detected as described (see text). Amino terminal capture or detection was determined using a variety of Ab1-16 antibodies; similar results were seen using 4G8 (directed against Ab17-24). Antibody 13.1.1 (top panel) is highly selective for Ab species ending at position 40, while antibody 2.1.3 (bottom panel) is highly selective for Ab species ending at position 42.
- Supplemental Figure 2 - Ab9 and Ab42-5 Antibody staining of frozen AD sections. Antibodies were diluted 1:100 and used to stain frozen cryostat sections of AD brains. Staining is shown for Ab9 (A, top panel) and Ab42-5 (B, bottom panel). Antibody staining was detected using a Goat anti-Mouse fluorescein-tagged antibody (Alexa Flor-488, Molecular Probes). Both dense and diffuse Ab plaques were readily stained by these antibodies.
- Supplemental Figure 3 - Microglia isolated from wild type mice readily internalize anti-Ab immune complexes into cytosolic compartments. Microglia isolated from (A) wild type mice and (B) FcRg-/- mice were incubated with Cy3-Ab complexed with Ab9 antibody (20 mg/ml) in the presence of fucoidan (500 mg/ml) for 10 min. Magnification 60x.
- Supplemental Figure 4 - Ab levels were only slightly reduced in the Ab1-42 immunized 14-15 month old Tg2576 x FcRg-/- mice. Mice were sacrificed following immunization with Ab1-42 for 3 months and both SDS soluble (SDS) and SDS-insoluble formic-acid extractable (FA) fractions were analyzed by capture ELISA. (A) 14-15 month old Tg2576 x FcRg-/- mice, n = 4/group); (B) 14-15 month old Tg2576 wt mice, n = 5/group). These changes were not significant, although a trend towards reduction was observed in the FA fractions of Ab42 in the Tg2576 x FcRg-/- group (Fig 2A, 28% reduction versus control group, P < 0.06).