Immunohistochemistry (Frozen sections) abreview for Anti-NeuN antibody [EPR12763] - Neuronal Marker

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Abreviews
Application Immunohistochemistry (Frozen sections)
Sample Human Cell (Pluripotent stem cell-derived retinal organoid)
Permeabilization Yes - 0.3% Triton X-100
Specification Pluripotent stem cell-derived retinal organoid
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 21°C
Fixative Paraformaldehyde

Other product details

Incubation time 16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 0.3% Triton X-100, 10% Serum, 0.1% BSA.
Dilution 1/500

Secondary antibody

Dilution 1/500
Name Non-Abcam antibody was used: Donkey anti-rabbit Alexa Fluor 546
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor­ 546

Additional data

Additional Notes Immunohistochemical analysis of NeuN-positive neurons. hPSC derived retinal organoids were washed once in PBS and then fixed in 4% paraformaldehyde (PFA) for 30 min, then immersed in 20% sucrose/PBS at 4C overnight. 20% sucrose/PBS was removed completely and the samples embedded in OCT matrix and frozen in liquid nitrogen. For immunohistochemistry, sections were blocked and permeabilised with PBS containing 0.3% triton X-100, 10% serum and 1% bovine serum albumin (block/permeabilization solution) for 1 hour. Abcam's NeuN primary antibody (white) was incubated overnight at 4C. Following primary antibody incubation, sections were incubated with Alexa-Fluor secondary antibody in 1:500 dilutions at room temperature for 2 hours, washed in PBS and counterstained with DAPI (Blue). Images were captured with a confocal microscope.
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Submitted Mar 29 2017