Figure 2
Characterization of immunoreactivity of 11A1 antibody. (A) Assessment of the binding affinity of 11A1 antibody against various synthetic Aβ peptides. E22P-Aβ42 (green), E22P-Aβ9-35ox (black), E22P-Aβ11-35ox (red), E22P-Aβ16-35ox (blue), and E22P-Aβ11-34 (orange) were synthesized and the binding affinity of 11A1 antibody for each peptide was assessed by Enzyme immunoassay. The 11A1 antibody exhibited high binding affinity for E22P-Aβ42 and E22P-Aβ9-35ox. Data are expressed as mean ± SD of three independent measurements. (B) Comparison of 11A1 antibody staining pattern with that of 6E10, 4G8, and OC antibodies. Postmortem human frontal cortex sections of an Alzheimer’s disease patient were stained with 6E10 (left panel), 4G8 (center left panel), OC (center right panel) and 11A1 (right panel) antibodies. Senile plaques (arrow heads) and vascular amyloid (arrows) are identified by 11A1 antibody with a similar pattern to that of the other three Aβ antibodies. The images were taken under 20x objective lens. Scale bar 50 µm. (C and D) Assessment of 11A1 cross-reactivity against APP. Histological sections of human cingulate gyrus with traumatic axonal injury were stained with 22C11 (APP antibody) (left panel) and 11A1 (right panel) antibodies. The traumatic axonal balloons display intense signal with the antibody for APP (arrowheads), but these lesions are not stained with 11A1 antibody. (C) The images were taken under 20x objective lens. Scale bar 50 µm. (D) The images were taken under 40x objective lens. Scale bar 20 µm. (E) Testing 11A1 antibody specificity for Aβ toxic conformer in senile plaques and blood vessels. 11A1 antibody was pre-incubated without (left panel) or with (right panel) its immunogen, E22P-Aβ9-35ox peptide, prior to incubation of tissues with the primary antibody. The pre-incubation with immunogen abrogates the signal in senile plaques and blood vessels. The images were taken under 20x objective lens. Scale bar 50 µm.
