NeuroMab™ Anti-S100A9 BBB Shuttle Antibody, Clone 6B4

Fig.1 monoclonal anti-S100A9 antibodies bind to the last 10 amino acids (peptide C7) of the C-terminal region of S100A9.

Representative micrographs of brains sections from 6 month old PSAPP mice treated with a single dose of engineered antibody (BL356) or placebo/control for 7 days. Sections were processed using S100B + TLR4 primary antibodies (brown; solid arrows) and counterstained with hematoxylin to visualize nuclei (blue; dashed arrows). Bars = 50 μm.

Fig.2 shows that the continuous epitope recognized by 6B4 is PGLGEGTP (C7a).

96 well high binding plate was coated with increasing concentrations of S100A9 peptide. After blocking the non-specific binding site with PBS/0.1% Tween/2% BSA, a solution containing 6B4 (1 μg/ml) was added to the wells and incubated for 1 hour at room temperature. As a negative control, some wells were incubated with PBS only (2e Ab). After extensive washing, HRP-conjugated goat anti-mouse was added to the plate and incubated for 1 hour at room temperature. The reaction was confirmed by addition of TMBS and stopped with H 2 SO 4. The optical density was read at 450 nm.

Fig.3 shows that humanized 6B4 recognizes the C-terminal peptide of S100A9.

Recombinant human (S100A9) mouse S100A9 (Mu14) (1 μg), PBS1x and C-terminal S100A9 peptide (2 μg) were put into a PVDF membrane, and a dot blot analysis was performed using humanized 6B4Hh + Lh or Hc + Lh.