Rabbit anti mtor antibody

For double-labelled immunofluorescence, 4-µm brain sections were deparaffinized in xylene, hydrated with a graded alcohol series, blocked with 5% non-immune fetal bovine serum (CAS: 9048-46-8; Sigma-Aldrich; Merck KGaA) in PBST (0.5% Tween-20 in PBS) at 37°C for 2 h and incubated with primary antibodies overnight at 4°C with rabbit anti-CB2R antibody (1:100; cat. no. ab3561; Abcam), rabbit anti-mTOR antibody (1:200; cat. no. 20657-1-AP; Proteintech, Inc.), rabbit anti-LC3B antibody (1:400; cat. no. 2775; Cell Signaling Technology, Inc.), and mouse anti-NeuN antibody (1:800; cat. no. MAB377; EMD Millipore). Afterward, an Alexa Fluor 594-conjugated donkey anti-rabbit IgG (1:200; cat. no. ab150076; Abcam) and an Alexa Fluor 488-conjugated donkey anti-mouse IgG (1:200; cat. no. ab150105; Abcam) was used as secondary antibodies conjugated with CB2R and NeuN, mTOR and NeuN, or LC3B and NeuN. After washing with PBS three times, the sections were covered with the VECTASHIELD Mounting Medium glasses. For the evaluation of CB2R⁺/NeuN⁺ cells, mTOR⁺/ NeuN⁺ cells and LC3⁺/NeuN⁺ cells, 10 microscope fields of the CA1, CA3, and DG regions were randomly selected under ×200 magnification by fluorescence light microscopy (Nikon Eclipse 80i; Nikon Corporation).