Rabbit anti goat antibody

After the behavioral test, the SNs of the rats were rapidly dissected out and homogenized in lysis buffer (Beyotime Inst. Biotech, Beijing, China). The microglial cells were collected and lysed with a lysis buffer. Supernatants were collected and the protein concentrations were measured using a bicinchoninic acid protein assay kit (Beyotime Inst. Biotech). A total of 30 µg of protein was subjected to 10% sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto immunoblot polyvinylidene difluoride membranes (Millipore, Billerica, MA, USA). After blocking with 5% nonfat milk, the blots were incubated overnight at 4 °C with primary antibodies against iNOS (1:2000), COX-2 (1:1000), OX-42 (1:1000), TH (1:1000) (Abcam), phosphor-ERK1/2 (1:2000), ERK1/2 (1:2000), phosphor-p38 (1:2000), p38 (1:1000), phosphor-JNK (1:1000), JNK (1:2000), phosphor-NF-κB p65 (1:1000), NF-κB p65 (1:1000) (Cell Signaling Technology, Danvers, MA, USA), and β-actin (1:2000) (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA). After this, the blots were incubated with a horseradish peroxidase-labeled secondary goat anti-rabbit (1:2000; Santa Cruz, CA, USA) or rabbit anti-goat antibody (1:2000; Santa Cruz, CA, USA) for 1 h at room temperature.

Caerulein (CAE) was purchased from Sigma-Aldrich (St. Louis, MO, United States). Rabbit monoclonal antibodies against OPA1 (ab157457, Abcam Plc, Cambridge, United Kingdom), DRP1 (D6C7 rabbit mAb; Cell Signaling), LC3B [LC3B antibody (2775)]; Cell Signaling Technology, MA, United States), p62 [SQSTM1 (P-15): sc-10117; Santa Cruz Biotechnology, Santa Cruz, CA, United States)], VMP1 (D1y3E, Cell Signaling Technology, MA, United States); mouse monoclonal antibodies against Parkin (P6248, Sigma-Aldrich, St. Louis, MO, United States), V5 [13202 V5-Tag (D3H8Q) rabbit mAB-Cell Signaling)], β-actin [β-actin (C4): sc-47778; Santa Cruz Biotechnology, Santa Cruz, CA, United States)], beta tubulin (ab131205, Abcam Plc, Cambridge, United Kingdom); rabbit anti-goat antibody (Santa Cruz Biotechnology, Santa Cruz, CA, United States); goat policlonal antibodies against VDAC-1 (D-16 sc-32063; Santa Cruz Biotechnology, Santa Cruz, CA, United States); rabbit anti-mouse antibody [(315-035-048) Jackson InmunoResearch, Baltimore Pike, United States]; goat anti-rabbit antibody [(GAR):170-5046; Bio-Rad, CA, United States]. Other reagents, enzymes, and chemicals were of reagent grade and also from Sigma-Aldrich.