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Love letter of great men book pdf free download. Love letter of great men book pdf free download.-amyloid plaque burden in the hippocephalus was similar to that of the spiny eel

Immunocytochemistry, analysis of Aβ and Congo red, and ELISA {#Sec8}
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Rabbits were deeply anesthetized using a mixture of Tricaine methanesulfonate (400 mg/ml) and Algavert^®^ (100 mg/ml; Sigma-Aldrich) and fixed by perfusion of freshly prepared 4% paraformaldehyde (Wako, Tokyo, Japan) at pH 7.4. Transverse hippocampal slices (400 µm thick) were obtained and fixed overnight in the same fixative at 4 °C and then washed three times with phosphate-buffered saline. They were then dehydrated in 30% sucrose at 4 °C. Whole brain samples were fixed with 4% paraformaldehyde overnight at 4 °C. All other steps were performed as previously described^[@CR11],\ [@CR12]^.

Using the rabbit anti-amyloid beta peptide \[1–16\] antibody (1:250; Abcam, Cambridge, UK) as the primary antibody, immunostaining was conducted at 4 °C overnight. The second antibody used was goat anti-rabbit IgG conjugated with Alexa Fluor 488 (1:1000, Life Technologies, Carlsbad, CA, USA). For Aβ immunostaining, sections were labeled with the oligomer-specific monoclonal antibody, clone OC (1:100; Thermo Fisher Scientific, Waltham, MA, USA). The confocal images were acquired using an A1RMP-SI microscope (Nikon, Tokyo, Japan).

Next, we estimated the total load of Aβ within the hippocampal CA3 subfield and other brain areas. We employed the clone OC, which recognizes oligomeric Aβ, and the rabbit anti-Aβ (1–16) antibody, which recognizes primarily monomeric Aβ. To reveal more detailed Aβ distribution, the clone OC antibody was applied for the following secondary immunostaining to identify
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