references
 
 
 

NeuN

NeuN expression is observed in most neuronal cell types throughout the adult nervous system. Immunohistochemically detectable NeuN protein first appears at developmental time points, which correspond with the withdrawal of the neuron from the cell cycle and/or with the initiation of terminal differentiation of the neuron (Mullen et al. 1992). This protein is useful and important in the determination of neuronal phenotypes within brain tissue.
   
 

NeuN Protocol

Positive Control Tissue: Rat hippocampus fixed with 4% paraformaldehyde.

Tissue: Rat Hippocampus

Fixation: 4% Paraformaldehyde in PBS pH 7.2

Primary Antibody: Mouse anti-NeuN

Ideal Dilution Ratio 1:500-1000

Secondary Antibody: Alexa 488/568 Goat anti-mouse IgG (H+L), (Molecular Probes, Cat# ).

Optimal dilution 1:200

  • Sections were washed with PBS to remove any sodium azide.
  • Sections were incubated with NeuN antibody. The Monoclonal NeuN antibody (Monoclonal, mouse anti-NeuN; Chemicon), was diluted at a ratio of 1:1000 in 0.3% Triton-X in PBS solution and incubated for 48 hours at 4 C with gentle shaking.
  • The sections were then washed 3 times for 5 minutes each in PBS.
  • Sections were then incubated in the dark for 2-hours with fluorescent-conjugated secondary antibody (Alexa 488/568 goat anti-mouse, 1:200 in 0.3% Triton-X solution; Chemicon) at room temperature.
  • Wash in distilled water 2x5min.
  • Coverslip with mounting medium (Permafluor for fluorecent tissues and Permount for DAB tissues).