Rabbit anti il6

Western blotting was done, following the previously mentioned method [18 ,19 (link)]. Hippocampal tissues were extracted by homogenization using lysis buffer (pH, 7.6) After separating 30 μg protein using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the protein was moved to a nitrocellulose membrane (GE Healthcare Bio-Sciences, Pittsburgh, PA, USA). The membrane was treated with 5% skimmed milk powder (BD Difco, Detroit, MI, USA) for blocking, and incubated with 0.1% Tween-20 (Sigma Aldrich Co.) for 1 hour at room temperature. The membrane was treated with the primary antibodies such as rabbit anti-VCAM-1 (1:1,000, Cell Signaling Technology), rabbit anti-ICAM-1 (1:1,000, Cell Signaling Technology), mouse anti-TNF-α (1:1,000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-IL-6 (1:1,000; Santa Cruz Biotechnology), and mouse anti-IL-1β (1:1,000; Santa Cruz Biotechnology). Membranes were subsequently incubated for l hour with peroxidase-tagged secondary antibodies. Enhanced chemiluminescence detection kit (DoGen, Seoul, Korea) was used for visualization of protein, and quantitative analysis of bands was calculated using Image-ProPlus program (Media Cybernetics, Rockville, MD, USA).