Fig. 3

FCA3340 specifically recognizes native and denaturated A β 40
(A) Two micrograms of Aβ40 was electrophoresed on a 16.5% Tris-tricine gel and Western blotted, and nitrocellulose sheets were incubated for 24 hr at 4°C with a 1:100 dilution of the IgG-purified fraction of FCA3340 (in TBS containing 5% skim milk) preincubated (for 24 hr at 4°C) without (lane C) or with 0.1 mM of synthetic octapeptides corresponding to Aβ33–40 (lane 1) or Aβ35–42 (lane 2); then immunological complexes were revealed by ECL as described in Materials and Methods. In B, 2 µg of Aβ were dot blotted, then hybridized for 18 hr at 4°C with a 1:100 dilution of FCA3340 preincubated for 24 hr at 4°C without (lane C) or with the synthetic peptides (0.1 mM) corresponding to Aβ33–40 (lane 1), Aβ35–42 (lane 2), or Aβ35–43 (lane 3). Immunoreactivities were revealed as above.