Rabbit anti il 6

Mouse brain tissue samples were collected for western blot analysis using 10–15% SDS-polyacrylamide gels to resolve protein samples. The gels were then electrophoretically transferred to 0.45-mm polyvinylidene difluoride membranes (Millipore, Billerica, MA, USA). After blocking with 8% milk in Tris-buffered saline with Tween (TBST) at room temperature for 1 h, rabbit anti-Iba-1 (1:1000; Wako, Japan), rabbit anti-IL-1β (1:500; Wanleibio, China), rabbit anti-IL-6(1:1000; Proteintech, China), mouse anti-GAPDH (1:10000; Proteintech, China) or mouse anti-α-tubulin (1:3000; Invitrogen, USA) primary antibodies were incubated with the membranes overnight at 4 °C. On the second day, the membranes were washed in TBST and then incubated with horseradish peroxidase-conjugated secondary antibodies for 1 h (1:5000; Proteintech, China). TBST was then used to wash the membrane three times (10 min each time). The membranes were placed on a gel imager, and enhanced chemiluminescence reagent (Thermo, Marina, CA, USA) was added to visualise the bands.