Mouse anti gfap antibody

After postfixation and incubation in gradient sucrose solutions, the L_3–5 segments of the spinal cord were sectioned into 30-μm frozen slices according to the method described above. The frozen slices were washed in PBS, and then high-pressure epitope retrieval (2 min) was performed in sodium citrate solution. The sections were incubated for 30 min in PBS containing 1% donkey serum and 0.5% Triton X-100 at 37°C and then incubated overnight at 4°C with a rabbit anti-p-p65 (Ser536) antibody (1:500, Cell Signaling Technology), a mouse anti-GFAP antibody (1:500, Cell Signaling Technology) a mouse anti-NeuN antibody (1:500, Abcam), or a goat anti-IBA-1 antibody (1:500, Abcam). The sections were rinsed in PBS three times for 15 min and then incubated for 30 min at 37°C with corresponding secondary antibodies (conjugated to Alexa FluorVR 488 and 594, Abcam). The samples were finally examined with a Fluorescence Inversion Microscope System (Leica, German), and images were acquired and processed using Leica software.