NeuroMab™ Anti-S100A9 Antibody(NRP-0422-P558)

Figure 1 Monoclonal anti-S100A9 antibody binds to the last 10 amino acids (peptide C7) of the C-terminal region of S100A9.

Representative photomicrographs of brain sections from 6-month-old PSAPP mice treated with a single dose of engineered antibody (BL356) or placebo/control for 7 days. Sections were subjected to proximity junction assay using S100B + TLR4 primary antibody (brown; solid arrow) and counterstained with hematoxylin to visualize nuclei (blue; dashed arrow). Bar = 50 μm.

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

96-well high-binding plates coated with increasing concentrations of the S100A9 peptide. After blocking non-specific binding sites 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 adding TMBS and quenched with H2SO4. Read the optical density at 450 nm.

Figure 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 plated on PVDF membranes and subjected to dot blot analysis using humanized 6B4Hh + Lh or Hc + Lh.