Rabbit anti syntaxin

Total proteins were extracted from the ischemic cortex 14 days after MCAO (n=3 for each group). Western blots were performed as previously described.23 (link) Equal amounts of sample from each group were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membranes. The membranes were blocked in 5% nonfat milk for 1 hour at room temperature and then incubated overnight at 4°C with the desired primary antibodies: rabbit anti-GADD45α (1:1,000; CST, Danvers, MA, USA), rabbit anti-Bcl-2 (1:1,000; CST), rabbit anti-Bax (1:1,000; CST), rabbit anti-KIFC2 (1:200; Santa Cruz Biotechnology Inc., Dallas, TX, USA), rabbit anti-SNAP25 (1:1,000; CST), rabbit anti-Syntaxin (1:5,000; Abcam, Cambridge, UK), rabbit anti-Complexin-1/2 (1:1,000; CST), rabbit anti-UCP3 (1:200; Novus, San Jose, CA, USA), and mouse anti-β-actin (1:1,000; Applygen, Beijing, People’s republic of China). After washing three times with tris-buffered saline-Tween 20 (TBST), the membranes were incubated with secondary antibodies conjugated to HRP for 1 hour. The immunoblots were visualized with enhanced chemiluminescence and analyzed using GelPro software. For each blot, β-actin served as an internal loading control.