Anti-NeuN 抗体 [1B7] - Neuronal Marker

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

IHC image of NeuN staining in rat brain 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 (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab104224, 1 μg/ml, for 15 minutes 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-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

IHC image of FOX3/NeuN staining in human normal hippocampus formalin fixed paraffin embedded tissue section performed on a Leica BondTM 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 minutes. The section was then incubated with ab104224 5 μg/ml for 15 minutes 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

Immunofluorescent analysis of rat brain stem co-stained with ab104224 in green and a chicken pAb to microtubule associated protein 2 (MAP2) in red. Blue is DAPI staining of nuclear DNA.

Following transcardial perfusion with 4% paraformaldehyde the brain was post fixed for 24 hours cut to 45μM and free-floating sections were stained. The Fox3/NeuN antibody selectively stains nuclei and the proximal cytoplasm of neuronal cells while the MAP2 antibody labels dendrites and overlaps with Fox3/NeuN staining in the perikarya of neurons.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

Rat brain neural cultures stained with ab104224 in pink with ab4674 (chicken polyclonal to GFAP) in green and DNA in blue. ab104224 reveals strong nuclear and distal cytoplasmic staining. It does not stain astrocytes and other non-neuronal cells.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

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 (pH 6 epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab104224 1 μg/ml for 15 minutes at room temperature. A goat anti-mouse 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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

ab104224 staining NeuN - Neuronal Marker in primary hippocampal rat neurons/glia (obtained from Neuromics cat. no. PC35101) DIV14. The cells were fixed with 100% methanol (5 min) permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab104224 at 0.1μg/ml and ab6046 Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117 Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080 Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a high-content analyser (Operetta CLS Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

Immunofluorescence staining of SOX9 using ab196450 in primary hippocampal mouse neurons/glia (obtained from Transnetyx Tissue by BrainBits LLC cat.no. C57EHP) DIV14. The cells were fixed with 4% formaldehyde (10 min) permeabilized with 0.1% TritonX-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab196450 at 1 μg/ml (shown in green), ab4674 (anti-GFAP) at 1/1000 dilution and ab104224 (anti-NeuN) at 1/1000 dilution. Cells were then incubated with ab150176 Goat Anti-Chicken IgY H&L (Alexa Fluor^® 594) preadsorbed (shown in red) and ab150119 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 647) (shown in purple) all secondary antibodies at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).
As expected most GFAP positive cells are also SOX9 positive while NeuN positive cells are SOX9 negative.
Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

Immunofluorescence staining of SOX9 using ab185966 in primary hippocampal mouse neurons/glia (obtained from Transnetyx Tissue by BrainBits LLC cat.no. C57EHP) DIV14. The cells were fixed with 4% formaldehyde (10 min) permeabilized with 0.1% TritonX-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab185966 at 1 μg/ml, ab4674 (anti-GFAP) at 1/1000 dilution and ab104224 (anti-NeuN) at 1/1000 dilution. Cells were then incubated with ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) ab150176 Goat Anti-Chicken IgY H&L (Alexa Fluor^® 594) preadsorbed (shown in red) and ab150119 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 647) (shown in purple) all secondary antibodies at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).
As expected most GFAP positive cells are also SOX9 positive while NeuN positive cells are SOX9 negative. SOX9 positive cells which are not GFAP positive (e.g. asterisk) are likely neural stem cells/ oligodendrocyte precursor cells present in the culture.
Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

Immunofluorescence staining of SOX9 using ab185230 in primary hippocampal mouse neurons/glia (obtained from Transnetyx Tissue by BrainBits LLC cat.no. C57EHP) DIV14. The cells were fixed with 4% formaldehyde (10 min) permeabilized with 0.1% TritonX-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab185230 at 5 μg/ml, ab4674 (anti-GFAP) at 1/1000 dilution and ab104224 (anti-NeuN) at 1/1000 dilution. Cells were then incubated with ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) ab150176 Goat Anti-Chicken IgY H&L (Alexa Fluor^® 594) preadsorbed (shown in red) and ab150119 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 647) (shown in purple) all secondary antibodies at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).
As expected most GFAP positive cells are also SOX9 positive while NeuN positive cells are SOX9 negative.
Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.

• WB

Supplier Data

Western blot - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

All lanes:

Western blot - Anti-NeuN antibody [1B7] - Neuronal Marker (ab104224) at 1/1000 dilution

Lane 2:

Adult rat whole brain lysate

Lane 3:

Embryonic E20 rat whole brain lysate

Lane 4:

Adult mouse whole brain lysate

Predicted band size: 34 kDa

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• IHC

CiteAb

Immunohistochemistry - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

Immunohistochemistry using Anti-NeuN antibody [1B7] - Neuronal Marker, ab104224. Publication image from Raynald et al., 2019, CNS Neurosci Ther, 31486601. Legend direct from paper.

A, The immunofluorescence assay of neurons in the area adjacent to the lesion area stained with NeuN (red) in longitudinal sections at 6 wk after SCI. The image (B) and quantification (C) of caspase3-positive staining within each group. D, Statistically significant differences in NeuN+ cells between the three groups. E, F, The mRNA expression of VEGFA (vascular endothelial growth factor A) and Casp 3 (caspase 3) showed a significant difference between three groups. **P < 0.01, *P < 0.05. Scale bar in (A) = 250 μm, scale bar in all magnification = 100 μm. Scale bar in (B) = 25 μm.

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

Immunocytochemistry-immunofluorescence using Anti-NeuN antibody [1B7] - Neuronal Marker, ab104224. Publication image from Tan, Z. et al., 2020, Ann Clin Transl Neurol, 32302063. Legend direct from paper.

The effect of FK866 on neuronal neurons in the peri-injured cerebral cortex at 24 h after TBI. In the rats subjected to TBI, the percentage of TUNEL-positive neurons was higher when compared with the sham group, while FK866 decreased neuronal apoptosis. Scar bar = 50 μm. n = 5/group. The bar represents mean ± SD, **P < 0.01 versus sham group; ##P < 0.01 versus TBI + vehicle group.

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

Immunocytochemistry-immunofluorescence using Anti-NeuN antibody [1B7] - Neuronal Marker, ab104224. Publication image from Fu, C. et al., 2020, Front Pharmacol, 32153398. Legend direct from paper.

Effect of QNDP on the expression of NLRP3, ASC, and cleaved caspase 1/pro-caspase 1 in MCAO rats. (A) Western blotting analysis of NLRP3, (B) Western blotting analysis of ASC, (C) Western blotting analysis of C-caspase 1/pro-caspase 1, (D) Immunofluorescent staining for NLRP3 (red), NeuN (green), the nuclei were stained blue with DAPI. Scale bar indicates 20 μm. Data are presented as the mean ± SEM, n = 3 per group, *p < 0.05, **p < 0.01 compared with the sham group, ##p < 0.01 compared with the MCAO group.

• IHC

CiteAb

Immunohistochemistry - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

Immunohistochemistry using Anti-NeuN antibody [1B7] - Neuronal Marker, ab104224. Publication image from Liu, D. et al., 2020, Front Pharmacol, 32194421. Legend direct from paper.

PUR alleviates HI-induced apoptosis at 72 h post-HI insult. (A) Levels of Bax and Bcl-2 within the ipsilateral cortex were measured with use of western blotting. Quantification of protein levels of Bax and Bcl-2 were determined with use of Image-Pro Plus 6.0 (N = 4/group). (B) Double immunofluorescent staining of active caspase-3 (red) and NeuN (green) within the ipsilateral cortex. Scale bar = 50 μm. (C) Quantification of active caspase-3/NeuN-positive cells (N = 4/group). Six images (20x) were captured randomly for each section per animal. (D) Immunoblotting analysis of active caspase-3, caspase-3 within the ipsilateral cortex. (E) Image-Pro Plus 6.0 was used to quantify protein levels of caspase-3 and active caspase-3. Results are expressed as the ratio of active caspase-3/caspase-3 (N = 4/group). Values represent the mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001 according to ANOVA with Bonferroni correction.

• IHC

CiteAb

Immunohistochemistry - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

Immunohistochemistry using Anti-NeuN antibody [1B7] - Neuronal Marker, ab104224. Publication image from Song, T. et al., 2022, Front Pharmacol, 35401208. Legend direct from paper.

The cellular localization of SIRT2 and FPN1 in the spinal cord. (A) SIRT2 levels decreased in the microglia of SNI rats. (B,C) FPN1 levels decreased in the microglia and neurons of SNI rats. The image in the first column on the left shows the overall outline of the spinal cord. Representative confocal images show the results of double immunofluorescence staining of SIRT2 or FPN1 (red) in the spinal dorsal horn; CD11b, a microglia marker (green) or NeuN, a neuronal marker (green) in the sham group and SNI group. Scale bars, 100 μm. SNI, spared nerve injury; SIRT2, sirtuin 2; FPN1, ferroportin 1.

• WB

CiteAb

Western blot - Anti-NeuN antibody [1B7] - Neuronal Marker (AB104224)

NeuN western blot using anti-NeuN antibody [1B7] - Neuronal Marker ab104224. Publication image and figure legend from Wang, J., Zhai, W., et al., 2018, Front Cell Neurosci, PubMed 29375318.

ab104224 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab104224 please see the product overview.

Protein levels and localization of DCC and UNC5H2 after ICH. (A) Western blot assay of protein levels of DCC and UNC5H2 in the peri-hematoma cortex at different time points after ICH. All values are expressed as mean ± SEM. Mean values of protein levels in the sham group were normalized to 1.0. *p < 0.05, **p < 0.01 vs. the sham group, n = 6. (B) Double immunofluorescence staining was performed in the peri-hematoma cortex with DCC antibody (green) and a neuronal marker (NeuN, red), and nuclei were labeled with DAPI (blue). Scale bar = 20 μm. All values are expressed as mean ± SEM. The mean value of the sham group was normalized to 1.0. **p < 0.01 vs. the sham group, n = 6. (C) Double immunofluorescence staining was performed in the peri-hematoma cortex with UNC5H2 antibody (green) and a neuronal marker (NeuN, red), and nuclei were labeled with DAPI (blue). Scale bar = 20 μm. All values are expressed as mean ± SEM. The mean value of the sham group was normalized to 1.0. **p < 0.01 vs. the sham group, n = 6.

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