Anti mao b

The brain tissues were homogenized with lysis buffer (PRO-PREP; iNtRON, Sungnam, Korea; n=8 mice per group) and centrifuged at 2500×g for 15 min at 4 °C. Equal amounts of total protein (20 μg) isolated from brain tissues were resolved on 8% or 10% sodium dodecyl sulfate polyacrylamide gels and then transferred to nitrocellulose membranes (Hybond ECL; Amersham Pharmacia Biotech, Piscataway, NJ). Membranes were incubated at 4 °C for 12 h with the following specific antibodies: anti-GFAP, anti-iba1 (1:1000; Abcam, Inc., Cambridge, MA), anti-COX-2, anti-p38, anti-p-p38, anti-p-JNK, anti-JNK, anti-p-ERK, anti-ERK (Cell Signaling Technology, Inc., Beverly, MA), anti-MAO B (1:1000; Santa Cruz Biotechnologies, Inc., Santa Cruz, CA), and anti-β-actin (1:2500; Santa Cruz Biotechnologies, Inc., Santa Cruz, CA). Blots were then incubated at room temperature for 2 h with corresponding peroxidase-conjugated anti-goat/mouse/rabbit (1/2000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA). Immunoreactive proteins were detected using an enhanced chemiluminescence [39] (link) Western blotting detection system. The relative density of the protein bands was scanned densitometrically using My Image (SLB, Seoul, Korea) and quantified by Lab Works 4.0 (UVP, Upland, CA).