200 kda anti neurofilament primary antibody

After fixation with 4 % paraformaldehyde for 20 min at room temperature (RT) and two washes with PBS (Gibco), the explants were permeabilized with 5 % triton X-100 (Sigma-Aldrich) for 10 min, washed twice with PBS, and blocked for non-specific antibody binding with 5 % donkey serum (Sigma-Aldrich). Neurites were labeled for neuro-filament using a mouse polyclonal 200 kDa anti-neurofilament primary antibody (1:400; Sigma-Aldrich). After primary antibody incubation overnight at 4 °C, followed by two PBS washes, the neurites were visualized by 2.5 h of incubation with fluorescein isothiocyanate (FITC)-conjugated secondary antibodies (1:100; Jackson Immunoresearch, West Grove, PA, USA) against mouse antibody. Specificity of staining was confirmed by a series of negative control staining without primary antibodies.

After fixation with 4% paraformaldehyde for 20 min at room temperature and two washes with PBS (Gibco), the explants were permeabilized with 5% triton X-100 (Sigma-Aldrich) for 10 min, washed twice with PBS and blocked for nonspecific antibody binding with 5% donkey serum (Sigma-Aldrich). Neurites were labeled for neurofilament using a mouse polyclonal 200 kDa anti-neurofilament primary antibody (1:400; Sigma-Aldrich). NrCAM stripes were visualized using polyclonal rabbit antibodies against NrCAM (1:400; R&D Systems). After primary antibody incubation overnight at 4°C, followed by two PBS washes, the neurites and stripes were visualized by 2.5h of incubation with fluorescein isothiocyanate (FITC) or Texas red (TR) conjugated secondary antibodies (1:100; Jackson Immunoresearch, West Grove, PA USA) against the species of the respective primary antibody. Specificity of staining was confirmed by a series of negative control staining without primary antibodies.