Anti aβ42 rabbit polyclonal

The immunogold procedure was done as we previously described (Sanchez-Varo et al., 2012 (link); Gomez-Arboledas et al., 2018 (link)). Sections (50 μm) from 4.5- and 6-month-old APP/PS1 mice were first washed in PBS and incubated in a 50 mM glycine solution 10 min, or cryoprotected in a 25% sucrose and 10% glycerol solution, followed by freezing at −80°C, to increase the antibody-binding efficiency. After following the standard protocol, the tissue was incubated overnight in primary anti-oligomeric OC rabbit polyclonal (1:5,000, Millipore), mouse monoclonal anti-oligomeric Aβ (12–24 mers) NU-4 (1:2,000), or anti-Aβ42 rabbit polyclonal (1:5,000, Abcam) antibodies in a PBS 0.1 M/1% BSA solution at room temperature. Then, sections were washed in PBS and incubated with 1.4 nm gold-conjugated secondary antibody (1:100, Nanoprobes) overnight at room temperature. After post-fixing with 1% glutaraldehyde and washing with 50 mM sodium citrate, the labeling was enhanced with the HQ Silver^TM Kit (Nanoprobes). Finally, the immunolabeled sections were processed as we previously described by the osmium fixation, dehydration, and embedding steps. The primary antibody was omitted in negative control experiments.

The following primary antibodies and dilutions were used: anti-Aβ42 rabbit polyclonal (1:5,000, Abcam); anti-Aβ (clone 6E10) mouse monoclonal (1:5,000, Signet); anti-human Aβ, N-terminus (clone 82E1) mouse monoclonal (1:6,000, IBL); anti-acetylated α-tubulin (clone 6-11B-1) mouse monoclonal (1:50,000, Sigma); anti-human amyloid precursor protein (hAPP) rabbit polyclonal (1:20,000, Sigma); anti-ATP synthase β (clone 10/ATP) mouse monoclonal (1:10,000, Brand BD Transduction Laboratories); anti-β-Actin (clone AC 74) mouse monoclonal (1:10,000, Sigma-Aldrich); anti-phospho-PHF-tau pSer202/Thr205 (clone AT8) mouse monoclonal (IHC 1:1,000, Cell Signaling); anti-PSD95 rabbit monoclonal (1:2,000, Cell signaling); anti-Somatostatin (SOM) goat polyclonal (1:1,000, Santa Cruz Biotechnology); anti-synaptophysin rabbit polyclonal (1:1,000, Abcam); anti-total tau (TAU46) mouse monoclonal (1:1,000, Cell Signaling) and anti-ubiquitin rabbit polyclonal (1:5,000, Dako).