Beta amyloid

A third cohort of mice was used for immunohistochemistry (see Figure 1). Sections were immunostained using standard free-floating bright field and fluorescent immunohistochemistry methods as previously described [29 (link), 32 (link)]. Briefly, all bright field sections were blocked with 80 mL PBS, 10 mL of methanol (Fisher Scientific), and 10 mL of 30% hydrogen peroxide (Fish Scientific) and incubated on a shaker (Model 55D, Reliable Scientific, Hernando, MS) for 15 min. Bright field and fluorescent sections were then washed 3 times and permeabilized for 30 min on a shaker with 1% Triton (Sigma-Aldrich, St. Louis, MO). Sections were incubated overnight with primary antibodies at room temperature, followed by a 2 h incubation with the appropriate secondary antibody at room temperature. The following primary antibodies were used with antibody identification number and working dilutions indicated in parentheses: Iba-1 (Invitrogen, Carlsbad, CA; AB_2544912, (1:1000)), GFAP (Agilent, Santa Clara, CA; AB_2811722, (1:10,000)), and beta-amyloid (Cell Signaling Technology, MA; AB_ 2797642, (1:500)).