Immunohistochemistry free floating abreview for Anti-NeuN antibody [EPR12763] - Neuronal Marker

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Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (brain)
Specification
brain

Other product details

Incubation time
24 hour(s) and 0 minute(s)
Dilution
1/2000

Secondary antibody

Dilution
1/500

Additional data

Additional Notes
Mice were perfused with 2% paraformaldehyde in PBS and postfixed overnight in 2% paraformaldehyde. Brains kept at 4 degrees in PBS until sectioning on a vibratome (60 um). Sections stored at -20 degrees in antifreeze until IHC reaction. Antigen retrieval with 1% sodium borohydride for 20 min. Pre-incubation in 10% normal goat serum, 1% BSA, 0.3% TritonX-100 in PBS for 1 hr. Primary incubation in 1% normal goat serum, 1% BSA, 0.3% TritonX-100, plus primary antibody (1:2000) in PBS overnight at room temp. Rinse 3 x 10 min in PBS. Secondary incubation in same solution as primary (but with secondary antibody) for 90 min at room temp. Rinse 3 x 10 min in PBS. Mount with Vectashield with DAPI. This antibody gave bright, low-background, specific labeling of neurons. Labeling intensity did decrease somewhat across the thickness of the tissue section, but it was still clearly visible in the center of the 60 micron section. In the image, green is GFP, red is NeuN, and blue is DAPI.

James Bogenpohl

Verified customer

Submitted Jul 19 2018

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