Immunohistochemical analysis of paraformaldehyde fixed frozen section Rat hippocampus tissue labeling NeuN with ab104225 at 1/1000 dilution.

IHC image of NeuN staining in rat cerebellum formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab104225, 1in500 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Immunofluorescent analysis of a section of adult mouse cerebellum stained with ab104225 at a dilution 1:5,000 in red, co-stained with chicken pAb to GFAP, dilution 1:5,000, in green.

Following transcardial perfusion of mouse with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45 μM, and free-floating sections. ab104225 stains the nuclei of neurons in the cerebellar granule layer. The GFAP antibody stains the processes of Bergmann glia in the molecular layer and astroglia in the granule and white matter layers.

IHC image of NeuN staining in human cerebellum formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab104225, 1in500 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

IHC image of NeuN staining in mouse cerebellum formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab104225, 1in500 dilution, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

IHC-P image of FOX3/NeuN staining using rat DRG using ab104225 (1:500). The sections were subjected to heat mediated antigen retrieval using citric acid pH 6. The sections were blocked using 1% BSA for 10 mins at 21°C. ab104225 was incubated for 2 hours at 21°C. The secondary antibody used was Goat polyclonal to Rabbit IgG conjugated to biotin (1:200).

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ab104225 at 1/1000 dilution, staining FOX3 (red) in rat brain cortex and counterstained for DNA in blue.

IHC-P image of FOX3/NeuN staining using mouse DRG using ab104225 (1:500). The sections were subjected to heat mediated antigen retrieval using citric acid pH 6. The sections were blocked using 1% BSA for 10 mins at 21°C. ab104225 was incubated for 2 hours at 21°C. The secondary antibody used was Goat polyclonal to Rabbit IgG conjugated to biotin (1:200).

See Abreview