Anti-NeuN antibody [EPR12763] - Neuronal Marker is a rabbit recombinant monoclonal antibody that is used to detect NeuN in Flow cytometry (Intra), ICC/IF, IHC (PFA fixed), IHC-Fr, IHC-P, Western blot, mIHC. Suitable for Cat, Common marmoset, Dog, Human, Mouse, Rat, Sheep, Zebrafish samples.
- Cited in over 865 publications
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Antibody clone EPR12763 is cited in over 1270 publications
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
WB | mIHC | ICC/IF | IHC-Fr | Flow Cyt (Intra) | IHC (PFA fixed) | IHC-P | |
---|---|---|---|---|---|---|---|
Human | Tested | Tested | Tested | Expected | Tested | Expected | Tested |
Mouse | Tested | Expected | Tested | Tested | Tested | Tested | Tested |
Rat | Tested | Expected | Expected | Expected | Expected | Expected | Tested |
Cat | Expected | Expected | Expected | Expected | Expected | Expected | Tested |
Common marmoset | Expected | Expected | Expected | Expected | Expected | Expected | Tested |
Cynomolgus monkey | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted |
Dog | Expected | Expected | Expected | Expected | Expected | Expected | Tested |
Goat | Expected | Expected | Expected | Expected | Expected | Expected | Tested |
Pig | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted |
Sheep | Expected | Expected | Expected | Expected | Expected | Expected | Tested |
Zebrafish | Expected | Expected | Expected | Expected | Expected | Expected | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 - 1/10000 | Notes For unpurified use at 1/1000 - 1/2000. |
Species Rat | Dilution info 1/1000 - 1/10000 | Notes For unpurified use at 1/1000 - 1/2000. |
Species Human | Dilution info 1/1000 - 1/10000 | Notes For unpurified use at 1/1000 - 1/2000. |
Species | Dilution info | Notes |
---|---|---|
Species Zebrafish, Common marmoset, Dog, Cat, Goat, Sheep | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Pig, Cynomolgus monkey | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 | Notes Perform Sodium citrate antigen retrieval (pH 6.0) in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Zebrafish, Common marmoset, Dog, Cat, Goat, Sheep | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Pig, Cynomolgus monkey | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/100 - 1/300 | Notes For unpurified use at 1/80. |
Species Human | Dilution info 1/100 - 1/300 | Notes For unpurified use at 1/80. |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Zebrafish, Common marmoset, Dog, Cat, Goat, Sheep | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Pig, Cynomolgus monkey | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Human, Zebrafish, Common marmoset, Dog, Cat, Goat, Sheep | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Pig, Cynomolgus monkey | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 | Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species Mouse | Dilution info 1/444 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Zebrafish, Common marmoset, Dog, Cat, Goat, Sheep | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Pig, Cynomolgus monkey | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Human, Zebrafish, Common marmoset, Dog, Cat, Goat, Sheep | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Pig, Cynomolgus monkey | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/3000 | Notes For unpurified use at 1/800. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/3000 | Notes For unpurified use at 1/800. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Zebrafish | Dilution info 1/3000 | Notes For unpurified use at 1/800. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Common marmoset | Dilution info 1/3000 | Notes For unpurified use at 1/800. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/3000 | Notes For unpurified use at 1/800. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Dog | Dilution info 1/3000 | Notes For unpurified use at 1/800. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Cat | Dilution info 1/3000 | Notes For unpurified use at 1/800. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Goat | Dilution info 1/3000 | Notes For unpurified use at 1/800. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Sheep | Dilution info 1/3000 | Notes For unpurified use at 1/800. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Pig, Cynomolgus monkey | Dilution info - | Notes - |
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Pre-mRNA alternative splicing regulator. Regulates alternative splicing of RBFOX2 to enhance the production of mRNA species that are targeted for nonsense-mediated decay (NMD).
RNA binding protein fox-1 homolog 3, Fox-1 homolog C, Neuronal nuclei antigen, NeuN antigen, RBFOX3
Anti-NeuN antibody [EPR12763] - Neuronal Marker is a rabbit recombinant monoclonal antibody that is used to detect NeuN in Flow cytometry (Intra), ICC/IF, IHC (PFA fixed), IHC-Fr, IHC-P, Western blot, mIHC. Suitable for Cat, Common marmoset, Dog, Human, Mouse, Rat, Sheep, Zebrafish samples.
- Cited in over 865 publications
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Antibody clone EPR12763 is cited in over 1270 publications
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Anti-NeuN antibody [EPR12763] - Neuronal Marker (ab177487) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC (PFA fixed), IHC-Fr, IHC-P, WB, mIHC in cat, common marmoset, dog, human, mouse, rat, sheep, zebrafish samples.
Anti-NeuN antibody (anti-RBFOX3 antibody) [EPR12763] - Neuronal Marker (ab177487) has been cited over 868 times in peer reviewed journals and is trusted by the scientific community.
Abcam's high quality manufacturing and validation processes ensure NeuN antigen antibody (ab177487) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
Anti-NeuN antibody [EPR12763] - Neuronal Marker (ab177487) has 72 independent reviews from customers.
Anti-NeuN antibody [EPR12763] - Neuronal Marker (ab177487) specifically detects NeuN (UniProt ID: A6NFN3; Molecular weight: 34kDa) and is sold in a convenient trial size to enable initial testing (20 µL) and larger sizes for subsequent scaling up experiments (100 µL and 1 mL).
Conjugation-ready, carrier free format available for antibody clone EPR12763 - Anti-NeuN antibody [EPR12763] - BSA and Azide free ab209898.
Antibody clone EPR12763 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor® 488, Alexa Fluor® 647, Biotin, Alexa Fluor® 594 (Alexa Fluor® 488 Anti-NeuN antibody [EPR12763] - Neuronal Marker ab190195, Alexa Fluor® 647 Anti-NeuN antibody [EPR12763] - Neuronal Marker ab190565, Biotin Anti-NeuN antibody [EPR12763] - Neuronal Marker ab204681, Alexa Fluor® 594 Anti-NeuN antibody [EPR12763] - Neuronal Marker ab207279).
NeuN (RBFOX3) is a protein extensively used as a neuronal marker due to its expression in the nuclei and cytoplasm of neurons. It is vital for the regulation of alternative splicing of pre-mRNA, which is essential for neural development and function. Mutations or dysregulation of RBFOX3 have been associated with various neurological conditions, including autism spectrum disorder, cognitive deficits, and epilepsy, highlighting its importance in maintaining normal brain function.
Browse our curated portfolio of extensively validated antibodies and assays, with antibodies to over 90% of key targets in major glioblastoma pathways– including EGF, VEGF, MAPK, Wnt, NFKB, and PI3K/AkT/MTOR – all in one place.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
NeuN also known as Fox-3 is a protein often utilized as a neuronal marker due to its expression predominantly in neurons. The molecular weight of NeuN is approximately 46-48 kDa. It is located in the nucleus and in the post-mitotic neurons of the central nervous system. Research frequently uses anti-NeuN antibodies for identifying neuronal cells in various samples capitalizing on its reliable expression profile across diverse neuron types. Common techniques for detecting NeuN include Western blot and immunohistochemistry (IHC) with NeuN staining being a definitive method for confirming neuronal identity.
NeuN plays a role in RNA splicing specific to neurons contributing to the regulation of gene expression within the nervous system. It does not function as part of a larger protein complex but interacts with chromatin to influence neuronal gene expression. This has implications for neuronal differentiation and maintenance. Techniques like NeuN IHC use antibodies such as Alexa Fluor 568 conjugated variants to visualize neurons in tissue confirming its utility as an accessible and specific neuronal marker in research.
NeuN serves an important function in neuronal development and differentiation. It particularly influences neurogenesis and synaptogenesis pathways where neuron-specific splicing and transcriptional regulation are essential. Although NeuN does not directly engage in pathways with other proteins because it operates independently its expression parallels the activity of significant neuronal proteins such as MAP2 and Neurofilaments. These relationships illustrate NeuN's indirect association in pathways promoting neuronal development and maintenance.
NeuN remains significant in studies of neurodegenerative diseases and neuronal damage. For instance decreased NeuN staining has been observed in Alzheimer's disease indicating loss of neuronal identity or survival. Research also notes alterations in NeuN expression following ischemic injury where its presence or absence links to neuronal viability. NeuN's association with these states of neuronal health or pathology highlights its relevance as a marker in neurodegenerative and injury-related disorders. Furthermore while not directly in disease pathways NeuN functionally intersects with proteins like Tau in Alzheimer's suggesting its indirect role in disease pathways.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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NeuN Immunohistochemistry (Frozen sections) staining of Mouse hippocampus (fresh frozen) using rabbit Anti-NeuN antibody
Immunohistochemical analysis of 4% PFA-fixed 0.2% Triton X-100 permeabilized frozen Mouse hippocampus (fresh frozen) tissue labeling MAP2 with Anti-MAP2 antibody [EPR19691] - Chicken IgY (Chimeric) ab318993 at 1/2000 (0.486 ug/ml) dilution followed by Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) ab150169 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) at 1/1000 (2 ug/mL) dilution (Green).
Panel A: merged staining of anti-MAP2 (Anti-MAP2 antibody [EPR19691] - Chicken IgY (Chimeric) ab318993 green), anti-NeuN (ab177487 red) and anti-GFAP (Alexa Fluor® 647 Anti-GFAP antibody [EPR1034Y] - Mouse IgG1 Chimeric ab313596 grey) on mouse hippocampus.
Panel B: anti-MAP2 stained on mouse hippocampus.
Panel C: anti-NeuN stained in neuron of mouse hippocampus.
Panel D: anti-GFAP stained in astrocytes of mouse hippocampus.
The nuclear counterstain was DAPI (Blue). The section was incubated with Anti-MAP2 antibody [EPR19691] - Chicken IgY (Chimeric) ab318993 ab177487 and Alexa Fluor® 647 Anti-GFAP antibody [EPR1034Y] - Mouse IgG1 Chimeric ab313596 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Secondary antibody control: Secondary antibody is Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) ab150169 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) at 1/1000 (2 ug/mL) dilution.
ab177487 staining NeuN in mouse free floating 50 micron lumbar spinal cord tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with formaldehyde, permeabilized with Triton X-100 and blocked with 10% serum for 2 hours at 25°C. Samples were incubated with primary antibody (1/500 in PBS + Triton) for 16 hours at 4°C. An Alexa Fluor® 594-conjugated donkey anti-rabbit IgG polyclonal (1/700) was used as the secondary antibody.
NeuN Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of mouse brain (frontal cortex) tissue using rabbit Anti-NeuN antibody
IHC-P image of FOX3/NeuN staining on mouse brain (frontal cortex) sections using ab177487 (1/800 dilution). Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/800 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250 dilution).
NeuN Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-NeuN antibody
IHC-P image of FOX3/NeuN staining on cat cerebellum sections using ab177487 (1/1000 dilution).
Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/1000 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250 dilution).
IHC-P image of FOX3/NeuN staining on zebrafish spinal cord sections using ab177487 (1/500). Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/500 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250).
NeuN Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of sheep brain tissue using rabbit Anti-NeuN antibody
IHC-P image of FOX3/NeuN staining on sheep brain (Frontal cortex) sections using ab177487 (1/1000 dilution). Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/1000 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250 dilution).
Fluorescence multiplex immunohistochemical analysis of human cerebellum tissue (formalin-fixed paraffin-embedded section).
Merged staining of Neu-N (ab177487; yellow; Opal™570), anti-beta III Tubulin (Anti-beta III Tubulin antibody [EP1569Y] - Neuronal Marker ab52623; red; Opal™690) and anti-GFAP (Anti-GFAP antibody [EPR1034Y] - Astrocyte Marker ab68428; green; Opal™520).
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ kit.
The section was incubated in three rounds of staining with ab177487 (1/1000 dilution), Anti-beta III Tubulin antibody [EP1569Y] - Neuronal Marker ab52623 (1/200 dilution) and Anti-GFAP antibody [EPR1034Y] - Astrocyte Marker ab68428 (1/250 dilution); each using a separate fluorescent tyramide signal amplification system.
Sodium citrate antigen retrieval (pH 6.0) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.
DAPI (blue) was used as a nuclear counter stain.
Immunofluorescence staining of NeuN using ab177487 in ioGlutamatergic Neurons (Human iPSC-Derived Glutamatergic Neurons, ioGlutamatergic Neurons - Human iPSC-Derived Glutamatergic Neurons ab259259), which were differentiated for 1 day post induction.
The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% PBS-Tween 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 ab177487 at 1 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cerebrum tissue labelling NeuN with ab177487 at 1/100 dilution (B), SOX1 with Anti-SOX1 antibody [EPR23041-60] ab242125 at 1/100 dilution (C) and Olig2 with Anti-Olig2 antibody [EPR2673] ab109186 at 1/100 dilution (D). Anti-Rabbit and Mouse Polymer HRP was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins. Heat mediated antigen retrieval (Leica ER2, PH9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibodies from the previous round, to avoid any cross-reactivity.
Panel A: merged staining of anti- NeuN (green, Opal™520), anti-SOX1 (red, Opal™570) and anti- Olig2 (yellow, Opal™690).
Panel B: anti-NeuN stained for neurons.
Panel C: anti-SOX1 stained on neural progenitors.
Panel D: anti-Olig2 stained on oligodendrocyte.
The section was incubated in three rounds of staining: in the order of ab177487, Anti-SOX1 antibody [EPR23041-60] ab242125 and Anti-Olig2 antibody [EPR2673] ab109186 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
NeuN Immunocytochemistry/ Immunofluorescence staining of Mouse primary neuron cells using rabbit Anti-NeuN antibody
Immunocytochemistry/immunofluorescence analysis of Mouse primary neuron cells labelling NeuN with ab177487 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 was used as the secondary antibody (green). Cells were counterstained with Anti-MAP2 mouse monoclonal antibody (Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267) at 1/200 dilution and visualised using Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/1000 dilution (red). Nuclear DNA was labelled with DAPI (blue).
Confocal image showing mainly nuclear staining in mouse primary neuron cells. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
NeuN Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-NeuN antibody
IHC-P image of NeuN (green) and GFAP (red) double staining on mouse cerebellum sections using ab177487 (1/5000) and Anti-GFAP antibody - Astrocyte Marker ab4674 (1/1500) respectively.
The sections were deparaffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were then incubated with Rabbit Monoclonal to NeuN (ab177487) diluted at 1/5000 and Chicken Polyclonal to GFAP (Anti-GFAP antibody - Astrocyte Marker ab4674) diluted at 1/1500. The primary antibody was detected using Goat Anti-Rabbit IgG Fc (Alexa Fluor® 488) preadsorbed ab150097 Goat anti-rabbit IgG conjugated to Alexa Fluor® 488 (1/500) and Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed ab150176 Goat anti-chicken IgY conjugated to Alexa Fluor® 594 (1/500)
Exposure time -
Lane 1-2: 3 minutes.
Lane 3-4: 1 minute.
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-NeuN antibody [EPR12763] - Neuronal Marker (ab177487) at 1/10000 dilution
Lane 1: Human fetal brain tissue lysate at 10 µg
Lane 2: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 10 µg
Lane 3: Mouse brain tissue lysate at 10 µg
Lane 4: Rat brain tissue lysate at 10 µg
All lanes: Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 34 kDa
Observed band size: 46 kDa, 48 kDa
NeuN antibody ab177487 was used with Tissue Clearing Kit Tissue Clearing Kit - hydrophobic ab243298 to penetrate, stain and clear a 1 mm coronal section of mouse brain. Blue: DAPI, Green: NeuN.
Learn more about tissue clearing kits, reagents, and protocols designed to make it easier to stain thick tissue sections and get more data from each valuable tissue section.
For 1 mm brain sections, we recommend a starting dilution of 1:200, and also using Goat Anti-Rabbit IgG H&L AlexaFluor488 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at a dilution of 1:400.
NeuN Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-NeuN antibody
Immunocytochemistry/immunofluorescence analysis of human neurons differentiated from iPSCs labelling NeuN (green) with ab177487 at 1/500 in 0.1% TritonX-100 1% goat serum 1X PBS for 16 hours at 4°C. Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100. Then cells were blocked with 5% serum for 20 minutes at 23°C. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilutionwas used as the secondary antibody. Tuj1 antibody was used to stain neuronal dendrites and axons (red).
Different batches of ab177487 were tested on mouse brain lysate at 2.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 46,48 kDa.
All lanes: Western blot - Anti-NeuN antibody [EPR12763] - Neuronal Marker (ab177487)
Predicted band size: 34 kDa
An independent comparison of commercially available NeuN clones in IHC-P.
Competitor A: Leading mouse monoclonal.
Competitor B: Non-Abcam rabbit monoclonal.
Sodium citrate was used for antigen retrieval in all 3 samples.
ab177487 produces specific staining, equivalent to the leading mouse monoclonal at half the dilution. The non-Abcam mouse monoclonal was less specific as it stained Purkinje cells, which do not express NeuN.
IHC image of NeuN (ab177487) with VHH Single Domain Anti-Rabbit IgG Fc (HRP) (VHH Single Domain Anti-Rabbit IgG Fc (HRP) ab191866) staining in formalin fixed paraffin embedded normal human cerebellum tissue section.
The section was dewaxed and then pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Dako Pascal pressure cooker using the standard factory-set regime. Non-specific protein-protein interactions were then blocked using in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1 hour at room temperature. The section was then incubated with rabbit monoclonal antibody [EPR12763] to NeuN (ab177487, 0.1μg/ml) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. Endogenous peroxidases were quenched using 1.6% (v/v) hydrogen peroxide in TBS containing 0.025% (v/v) Triton X-100 for 30 minutes at room temperature, with agitation. The secondary antibody, VHH Single Domain Anti-Rabbit IgG Fc (HRP) (VHH Single Domain Anti-Rabbit IgG Fc (HRP) ab191866, 1.0μg/ml) was then applied for 1 hour at room temperature in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA before being developed for 10 minutes at room temperature using Steady DAB/Plus (ab103723). The section was then counterstained with hematoxylin and mounted with DPX.
The negative control (secondary antibody only, no primary) inset shows no staining, demonstrating secondary antibody specificity.
For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.
NeuN Flow Cytometry (Intracellular) staining of U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells using rabbit Anti-NeuN antibody
Overlay histogram showing U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells stained with ab177487 (red line).
The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab177487 1/100 dilution) for 30 minutes at 22°C. The secondary antibody used was Alexa Fluor®488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 1μg/1x106cells used under the same conditions. Unlabeled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Alexa Fluor® 488 (Alexa Fluor® 488 Anti-NeuN antibody [EPR12763] - Neuronal Marker ab190195) and Alexa Fluor® 647 (Alexa Fluor® 647 Anti-NeuN antibody [EPR12763] - Neuronal Marker ab190565) conjugated versions are available for this clone.
NeuN Immunocytochemistry/ Immunofluorescence staining of SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells using rabbit Anti-NeuN antibody
Immunocytochemsitry/Immunofluorescence analysis of SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling NeuN (green) with ab177487 at 1/300 dilution. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200 dilution) was used as the secondary antibody. Counterstained with DAPI (blue).
An independent comparison of commercially available NeuN clones in IHC-Fr (acetone-fixed mouse dentate gyrus sections).
Competitor A: Leading mouse monoclonal.
Competitor B: Non-Abcam rabbit monoclonal.
ab177487 produces intense, specific staining with minimal background, even at half the dilution of competing antibodies.
NeuN Immunohistochemistry (Frozen sections) staining using rabbit Anti-NeuN antibody
ab177487 staining NeuN in mouse brain tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with formaldehyde and blocked with Triton X-100 + 0.4% horse serum for 30 minutes at 20°C. Samples were incubated with primary antibody (1/500 in blocking solution) for 16 hours at 4°C. An Alexa Fluor® 594-conjugated donkey anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.
NeuN Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of dog cerebellum tissue using rabbit Anti-NeuN antibody
IHC-P image of FOX3/NeuN staining on dog cerebellum sections using ab177487 (1/500 dilution).
Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/500 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250 dilution).
NeuN Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-NeuN antibody
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gliocytoma tissue labelling NeuN with ab177487 at 1/3000 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.
IHC-P image of FOX3/NeuN staining on rat brain (SVZ) sections using ab177487 (1/2000). Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/2000 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250).
IHC-P image of FOX3/NeuN staining on marmoset cerebellum sections using ab177487 (1/2000). Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/2000 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250).
NeuN Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of goat cerebellum tissue using rabbit Anti-NeuN antibody
IHC-P image of FOX3/NeuN staining on goat cerebellum sections using ab177487 (1/500 dilution). Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/500 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250 dilution).
Chromogenic multiplex immunohistochemical staining of FFPE normal human cerebellum tissue. ab177487, anti-NeuN DAB chromogen. Anti-GFAP antibody [EPR1034Y] - Astrocyte Marker ab68428, anti-GFAP purple chromogen and Anti-Iba1 antibody [EPR16588] ab178846, anti- Iba1 teal chromogen plus haematoxylin counterstain.
Chromogenic immunostaining was performed on a Roche Ventana Discovery Ultra instrument. The section was deparaffinised and incubated with CC1 solution for 24min 100°C. Following this with 3 rounds of staining in the order of ab177487 (1/600), Anti-Iba1 antibody [EPR16588] ab178846 (1/4000) Anti-GFAP antibody [EPR1034Y] - Astrocyte Marker ab68428 (1/1000). Between rounds of staining, antibody denaturation was conducted using Ultra CC2 solution for 8min at 100°C to avoid cross reactivity. Signal was developed with anti-rabbit HQ followed by anti-HQ HRP coupled with Chromomap DAB kit, Discovery purple or Discovery teal chromogens and haematoxylin II counterstain.
NeuN Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of human cerebellum tissue using rabbit Anti-NeuN antibody
Immunohistochemical analysis of formalin fixed paraffin embedded human cerebellum labelling NeuN with ab177487 at a dilution of 1/500. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an ChromoMap DAB (RUO) IHC Detection Kit with anti rabbit HQ and anti HQ HRP. Heat mediated antigen retrieval was conducted for 24 min with DISCOVERY cell conditioning solution (CC1) 100°C, pH 8.5. ab177487 was incubated at 37°C for 16 min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
NeuN Immunohistochemistry (Frozen sections) staining of Rat hippocampus (fresh frozen) using rabbit Anti-NeuN antibody
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat hippocampus (fresh frozen) tissue labeling MAP2 with Anti-MAP2 antibody [EPR19691] - Chicken IgY (Chimeric) ab318993 at 1/2000 (0.486 ug/ml) dilution followed by Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) ab150169 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) at 1/1000 2 ug/mL dilution (Green).
Panel A: merged staining of anti-MAP2 (Anti-MAP2 antibody [EPR19691] - Chicken IgY (Chimeric) ab318993, green), anti-NeuN (ab177487, red) and anti-GFAP (Alexa Fluor® 647 Anti-GFAP antibody [EPR1034Y] - Mouse IgG1 Chimeric ab313596, grey) on rat hippocampus.
Panel B: anti-MAP2 stained on rat hippocampus.
Panel C: anti-NeuN stained in neuron of rat hippocampus.
Panel D: anti-GFAP stained in astrocytes of rat hippocampus.
The nuclear counterstain was DAPI (Blue). The section was incubated with Anti-MAP2 antibody [EPR19691] - Chicken IgY (Chimeric) ab318993, ab177487 and Alexa Fluor® 647 Anti-GFAP antibody [EPR1034Y] - Mouse IgG1 Chimeric ab313596 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Secondary antibody control: Secondary antibody is Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) ab150169 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488)at 1/1000 2 ug/mL dilution.
Flow cytometry overlay histogram showing Neuro-2a cells stained with ab177487 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab177487) (1x 106in 100µl at 5.0µg/ml (1/444)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
This antibody gave a positive signal in Neuro-2a Fixed with 80% methanol (5 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.
NeuN IHC-tissue clearing staining of whole mouse brain using rabbit Anti-NeuN antibody
Anti-NeuN ab177487 was used with Tissue clearing kit – CUBIC (Tissue Clearing Kit – CUBIC ab316246) and 3D Tissue Staining Kit – CUBIC (3D Tissue Staining Kit – CUBIC ab316248) to penetrate stain and clear a whole mouse brain.
White: nuclear staining
Red: NeuN.
Learn more about tissue clearing kits reagents and protocols designed to make it easier to stain whole brains and get more data from each valuable tissue sample. For a whole mouse brain we recommend starting with 10 ug of ab177487 and using a Fab fragment secondary antibody with 6.67 µg to create an antibody complex before 3D staining (see protocol for details). Additive A was used during the staining process.
The sample was imaged using a light-sheet microscope.
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