Mouse monoclonal anti drp1 antibody

Immunohistochemical or immunocytochemical staining for 7 µm wax sections of ONHs or cultured ONH astrocytes were performed as described previously (Ju et al., 2008 (link)). Five sections per wax block from each group (n = 4 ONHs/group) were used for immunohistochemical analysis. The primary antibodies included mouse monoclonal anti-glial fibrillary acidic protein (GFAP) antibody (1:300; Sigma), guinea pig polyclonal anti-GFAP antibody (1:500; Advanced ImmunoChemical. Long Beach, CA), mouse monoclonal antineurofilament antibody (1:500; Sigma), mouse monoclonal anti-NR1 antibody (1:1,000; BD Pharmingen, San Diego, CA), rabbit monoclonal anti-NR2A antibody (1:100; Millipore, Billerica, MA), rabbit polyclonal anti-NR2B antibody (1:5,000; Millipore), and mouse monoclonal anti-DRP1 antibody (1:1,000; BD Transduction Laboratories, San Diego, CA). The images were acquired with a FluoView1000 confocal microscope (Olympus, Tokyo, Japan).

Cultured human ONH astrocytes were lysed with lysis buffer as described previously (Noh et al., 2013 (link)). The primary antibodies included rabbit polyclonal anti-NR1 and NR2A antibody (1:1,000; Cell Signaling Technology, Danvers, MA), rabbit polyclonal anti-GLAST (EAAT1, 1:500; Santa Cruz Biotechnology, Santa Cruz, CA), mouse monoclonal anti-DRP1 antibody (1:1,000; BD Transduction Laboratories), rabbit polyclonal anti-phospho-DRP1 at Ser616 antibody (1:1,000; Cell Signaling), and mouse monoclonal anti-actin antibody (1:5,000; Millipore). The scanned film images were analyzed with ImageJ (National Institute of Health, MD) and band densities were normalized to the band densities for actin.