Rabbit polyclonal anti bdnf

Hippocampi were dissected from animal after the end of LBP and SCO administration. Protein was extracted using a mammalian Protein Extraction Reagent (Pierce, Rockford, IL, USA) according to the manufacturer’s instructions and separated on 10% polyacrylamide gels in the presence of sodium dodecyl sulfate (SDS). Membranes after transferring were subsequently blocked with 5% non-fat milk in TBS and incubated with mouse monoclonal anti-IGF-1 (1∶1,000, Sigma, St Louis, MO), rabbit polyclonal anti-BDNF (1∶200), -Bax (1∶500), -Bcl-2 (1∶500, Santa Cruz, CA, USA) and mouse monoclonal anti-GAPDH (1∶10,000, Sigma, St Louis, MO, USA) overnight at 4°C. Membranes were developed by incubation with horseradish peroxidase-conjugated goat anti-mouse or rabbit IgG (1∶3,000, Pierce, Rockford, IL, USA) for 2 h at room temperature. After washing, the complexes were visualized by enhanced chemiluminescence (Santa Cruz, CA, USA) and exposed to X-ray film (Kodak, Rochester, NY, USA). The intensity of each band was quantified using the Shine-tech Image System (Shanghai, CHN).

Mouse monoclonal anti-NFκB p65, mouse monoclonal anti-CaMKII, rabbit polyclonal anti-BDNF, and rabbit polyclonal anti-integrin αM antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Mouse monoclonal anti-actin, mouse monoclonal anti-NR2B and rabbit polyclonal anti-NR2A antibodies were purchased from Millipore (Billerica, MA, USA). Mouse monoclonal anti-GFAP, rabbit monoclonal anti-phospho-NFκB p65, rabbit monoclonal anti-TNF-α, rabbit monoclonal anti-IL-6, mouse monoclonal anti-IL-1β, and horseradish peroxidase (HRP)-conjugated IgG antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). ECL Western Blotting Reagents were purchased from GE Healthcare (Little Chalfont, Buckinghamshire, UK). Other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA).