Rabbit anti bdnf antibody

According to a previously described method [13 (link),21 (link)], western blot was conducted. From the peri-infarct cortex and hippocampus, total protein was extracted using lysis buffer containing 150mM NaCl, 1mM phenylmethylsulfonyl fluoride, 1% Nonidet P40, 0.5% deoxycholic acid, 0.1% sodium dodecyl sulfate (SDS), 100-mg/mL leupeptin, 50mM Tris-HCl (pH, 7.5). Protein was quantified using a Bradford protein assay (Bio-Rad, Hercules, CA, USA). Protein, 30 µg, was separated on SDS polyacrylamide gel electrophoresis. After electrophoresis, proteins were transferred to nitrocellulose membrane (GE Healthcare Life Sciences, Chicago, IL, USA). The membrane was blocked with skim milk, then membrane was incubated by mouse anti-β-actin antibody (1:1,000; Santa Cruz Biotechnology), rabbit anti-PSD-95 antibody (1:1,000; Abcam, Cambridge, UK), rabbit antisynaptophysin (1:1,000; Abcam), rabbit antiBDNF antibody (1:1,000; Alomone Labs, Jerusalem, Israel), and rabbit anti-TrkB antibody (Santa Cruz Biotechnology) at 4°C during overnight. After washing, species appropriate horseradish peroxidase-conjugated secondary antibodies were incubated for 1 hour. Band detection was performed using the enhanced chemiluminescence detection system (Santa Cruz Biotechnology).