Monoclonal rabbit anti phospho gsk 3β ser9 p gsk 3β

We used a rabbit anti-Iba-1 (Wako Pure Chemical Industries Ltd., Osaka, Japan) antibody to immunohistochemically stain the ramified microglia and macrophages of rat spinal cords, and for Western blotting. We employed monoclonal rabbit anti-phospho-GSK-3β (Ser9) (p-GSK-3β) and monoclonal rabbit anti-GSK-3β antibodies (Cell Signaling Technology, Beverly, MA, USA) to detect GSK-3β. Rabbit polyclonal anti-Nrf-2 and -HO-1 antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA) were used to stain the respective proteins. We also employed a mouse monoclonal anti–β-actin antibody (Sigma-Aldrich, St. Louis, MO, USA).

For the immunohistochemical analysis of rat spinal cords, we used mouse monoclonal anti–glial fibrillary acidic protein (Sigma-Aldrich, St. Louis, MO, USA) for astrocytes and rabbit anti–ionized calcium-binding protein-1 (Iba-1) (Wako Pure Chemical Industries, Ltd., Osaka, Japan) for ramified microglia and macrophages.
To detect GSK-3β, monoclonal rabbit anti-phospho-GSK-3β (Ser⁹) (p-GSK-3β) and monoclonal rabbit anti-GSK-3β antibodies were used (Cell Signaling Technology, Beverly, MA, USA). Monoclonal rabbit anti-β-catenin (Ser⁶⁷⁵) and PhosphoPlus Akt (Ser⁴⁷³) antibody kits were purchased from Cell Signaling Technology. Rabbit polyclonal anti–vascular cell adhesion molecule-1 (VCAM-1) antibody (H-276, Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used to assess the severity of inflammation. A mouse monoclonal anti-β-actin (Sigma-Aldrich) antibody was also used.