Gapdh rabbit monoclonal antibody

Immunoblotting and quantification were performed as previously described (Pannasch et al., 2014). Hippocampi were collected in a small volume of cold SDS 2% containing a cocktail of protease inhibitors and phosphatase inhibitors (β‐glycerophosphate (10 mM) and orthovanadate (1 mM)) to which Laemmli 5× buffer was added. Samples were sonicated, boiled 5 min and loaded on 4–12% polyacrylamide gels. Equal amounts of proteins were separated by electrophoresis and transferred onto nitrocellulose membranes. Membranes were saturated with 5% fat‐free dried milk in triphosphate buffer solution and incubated overnight at 4 °C with primary antibodies (GAPDH rabbit monoclonal antibody [Sigma], Cx30 rabbit polyclonal antibody). They were then washed and exposed to peroxidase‐conjugated secondary antibodies (donkey anti‐rabbit IgG HRP‐conjugated secondary antibodies, Amersham Biosciences). GAPDH was used as loading control. Specific signals were revealed with the chemiluminescence detection kit (ECL, GE Healthcare). Semi‐quantitative densitometric analysis was performed after scanning the bands with the imageJ software.

Cortical tissue surrounding the subdural electrode (Fig. 5A) was sampled and homogenized in 2% SDS with protease inhibitor mixture, β-glycerophosphate (10 mM), and orthovanadate (1 mM). Equal amounts of protein were separated on a 10% PAGE gel followed by transfer to nitrocellulose membranes. Proteins were detected by immunoblotting using the HRP-ECL kit from Perkin-Elmer. GAPDH was used as loading control. Primary antibodies used were: GAPDH rabbit monoclonal antibody (Sigma), GFAP rabbit polyclonal antibody. Donkey anti-rabbit IgG (Amersham Biosciences) HRP-conjugated secondary antibodies were used.