Immunofluorescence Analysis of BDNF and GFAP

Immunofluorescence was then studied. The TG iced-slices were washed in PBS and incubated in a blocking solution containing 3% bovine serum albumin (BSA) in PBS with 0.3% Triton X-100 (Solarbio Company, Beijing, China) for 30 minutes at room temperature. In 4 °C refrigerator primary antibodies (mouse anti-GFAP, Abcam Company, 1:200; rabbit anti-BDNF, Abcam, 1:500) were incubated overnight. The samples were washed using PBS and the secondary antibody (goat anti-rabbit TRITC, Jackson ImmunoResearch Inc., West Grove, PA, USA, 1:200; goat anti-mouse FITC, Beijing Zhongshan Biotech Co., 1:200) was added for 45 minutes at 37 °C. Controls omitted the primary antibody. A fluorescence microscope (Olympus, Japan) and Image-Pro Plus 6.0 software were used to detect the immunofluorescence intensity of BDNF or GFAP expression. The co-expression steps of TrkB and GFAP were the same as mentioned above, except for the different dilution ratio (TrkB 1:1000, GFAP 1:200)^{27 (link)}.