Goat anti mouse igg

Proteins were extracted from cardiac fibroblasts or mouse myocardium by RIPA lysis buffer (Applygen, China) containing protease inhibitor cocktail (Sigma, Santa Clara, CA, USA). Total protein concentrations were measured by BCA assay (Pierce, USA). Proteins were then separated by SDS-PAGE and transferred to PVDF membranes (Merck Millipore, Germany). Membranes were incubated with primary antibodies collagen type I, Fibronectin (FN), Sec61α (Abcam, Cambridge, MA), TGF-β1, a-smooth muscle actin (α-SMA), connective tissue growth factor (CTGF) (Santa Cruz Biotechnology, CA, USA), α-Tubulin (Bioworld, USA), or Nogo-C (Abmart, China) and then incubated with secondary antibodies goat anti-rabbit IgG, goat anti-mouse IgG, rabbit anti-goat IgG (Biodragon, China), or mouse anti-rabbit IgG LCS (Abbkine, California, USA). Immunoblots were evaluated using the Chemi Doc XRS + instrument (Bio-RAD, Hercules, CA, USA).

Total protein was extracted from Schwann cells or mouse sciatic nerve with RIPA lysis buffer (Thermo Fisher Scientific) containing protease inhibitor cocktail (Sigma). Total protein concentration was measured using the BCA assay (Pierce, Rockford, IL, United States). Total protein was separated by SDS-PAGE and transferred to a PVDF membrane (Merck Millipore). The membrane was incubated with anti-Nogo-C (Abmart, Shanghai, China), β-actin, Krox-20 (Abcam, MA), and c-Jun, GFAP bcl2 or BAX (CST, United States) primary antibodies. Next, goat anti-rabbit IgG, goat anti-mouse IgG (Biodragon, Beijing, China), or mouse anti-rabbit IgG LCS (Abbkine, Wuhan, China) secondary antibodies were incubated. Immunoblotting was assessed using the Chemi Doc XRS + imaging system (Bio-Rad, Redmond WA, United States).