Ab77373

Western blot was used to detect the protein expression in tissues and cell lysates as previously described.20 (link),21 (link) The primary antibody were incubated in 4°C overnight and the primary antibody were anti-IL-12 p40 antibody (1:1000, ab77373, abcam, UK), anti-JAK2 antibody (1:500, ab39636, abcam, UK), anti-JAK2 (phospho Y1007) antibody (1:500, ab195055, abcam, UK), anti-STAT4 antibody (1:2000, ab235946, abcam, UK) and anti-STAT4 (phospho Y693) antibody (1:500, ab28815, abcam, UK). The second antibody was incubated for 1 hr at room temperature. BeyoECL Plus kit (P0018S, Beyotime, Shanghai, China) was used to chromogenic and densitometry protein bands with Beckman Coulter Immunoassay System (UniCel DxI 800, Beckman, CA, USA).

The brain tissue sections were fixed with 4% paraformaldehyde, 0.2% TritonX-100 was ruptured; Rabbit serum and rat serum were incubated for 1 hr at room temperature, and washed with PBS for 3 times; The primary antibody was incubated overnight at 4 degrees and the secondary antibody was incubated for 1 hr at room temperature. Primary antibodies: Anti-CD11b antibody (1:200, ab8878, abcam, UK) and anti-IL-12 p40 antibody (1:2000, ab77373, abcam, UK); Secondary antibodies: TRITC-conjugated goat-rabbit IgG (C5872, Jackson Immunoresearch, USA), Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (A11008, Invitrogen, USA). At last, 5 ug/mL DAPI (D8417, SIGMA, USA) at room temperature for 5 mins.