Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-NeuN [EPR12763] antibody - Neuronal Marker

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Abreviews
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step Serum as blocking agent for 15 minute(s) · Concentration: 10% · Temperature: 22°C
Antigen retrieval step None
Sample Mouse Tissue sections (Brain)
Specification Brain
Permeabilization Yes - 0.5% Triton X
Fixative Paraformaldehyde

Other product details

Incubation time 48 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1% Normal horse serum+0.1%Kodak photoflo
Dilution 1/1000

Secondary antibody

Name Non-Abcam antibody was used: Biotinylated Donkey anti Rabbit IgG (H+L)
Host species: Donkey
Clonality: Polyclonal
Conjugation: Biotin
Dilution 1/200

Additional data

Additional Notes Cornal section through the hippocampus showing NeuN positive neurons. 4% Paraformaldehyde was used to perfuse and fix the brain tissue followed by 20% sucrose in PBS over night. Sections were cut in cryostat at 25um. Tissue was incubated 0.5%Triton-X for permebalization for 30 min and blocked in 10% normal horse serum for 15 min and then in NeuN antibody at 1:1000 in antibody diluent that contains 1%horse serum serum+0.01 kodak photpflo for 48 h at 4C. Subsequently, tissue was incubated in Biotinylated Donkey anti-Rabbit IgG at 1:200 dilution. Tissue sections were then incubated in ABC solution (1:100) for 1 hr followed by DAB reaction to visulaize the immunoreaction product. The brown reaction product inidcates the positive reaction.
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Sumit Sarkar

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Submitted May 21 2014