FIGURE 5.
Immunoprecipitation of the dynein-dynactin complex and mutant SOD1 from brains and spinal cords of end-stage SOD1G93A mice using two different IP buffers. Homogenates of brains and spinal cords (SC) from three end-stage transgenic mice (SC-1–3) prepared in PBS without Ca2+ and Mg2+ (A) or in RIPA buffer containing 50 mm Tris-HCl (pH 7.4), 150 mm NaCl, 0.25% deoxycholic acid, 1% Nonidet P-40, 1 mm EDTA (B) were incubated with anti-DIC, anti-SOD1 antibody, or with IgG as a negative control, and the IP procedure was conducted as described by Zhang et al. (22). The IP products along with 2% of the input were analyzed by SDS-PAGE and Western blotting.