LC3B Antibody

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Publication protocol

Twenty-four hours after ISO, mice were perfused with PBS followed by PBS containing 4% paraformaldehyde. Brains were postfixed in 4% paraformaldehyde overnight. Forty-micron-thick coronal sections were cut with a vibratome. Sections were blocked in 5% normal goat serum, 1% BSA, 0.2% Triton X-100, 0.03% H2O2 in PBS for 1 h at room temperature and incubated with anti-LC3-antibody (1:800; Novus; NB600-1384) for 48 h at 4 °C; after washing with PBS, they were incubated with biotinylated anti-rabbit secondary antibody (1:200; Jackson ImmunoResearch; 711-065-152) for 2 h at room temperature, washed, and incubated with ABC reagent (Vector ABC kit; PK-6100) for 90 min at room temperature. After washing, sections were stained with diaminobenzidin (Vector Laboratories; SK-4100) for 1–2 min, washed, and sealed with a coverslip (21).

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