Recombinant
RabMAb

Anti-NeuN antibody [EPR12763] - Neuronal Marker (ab177487)

Overview

  • Product nameAnti-NeuN antibody [EPR12763] - Neuronal Marker
    See all NeuN primary antibodies
  • Description
    Rabbit monoclonal [EPR12763] to NeuN - Neuronal Marker
  • Tested applicationsSuitable for: IHC-FoFr, Flow Cyt, IHC-Fr, IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Goat, Cat, Dog, Human, Pig, Zebrafish, Cynomolgus monkey, Common marmoset
  • Immunogen

    Synthetic peptide within Human NeuN aa 1-100 (Cysteine residue). The exact sequence is proprietary.
    Database link: A6NFN3

  • Positive control
    • WB: Mouse brain, mouse cerebellum, rat cerebellum and human fetal brain tissue lysates. ICC/IF: SH-SY-5Y cells. IHC-P: Human cerebellum, human gliocytoma tissue. IHC-Fr: Mouse dentate gyrus tissue. Flow Cyt: U-87MG cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Alternative versions available:

    Anti-NeuN antibody (BSA & Azide free) [EPR12763] - Neuronal Marker (ab209898)

    Anti-NeuN antibody (Alexa Fluor® 488) [EPR12763] - Neuronal Marker (ab190195)

    Anti-NeuN antibody (Alexa Fluor® 647) [EPR12763] - Neuronal Marker (ab190565)

    Anti-NeuN antibody (Alexa Fluor® 568) [EPR12763] - Neuronal Marker (ab207282)

    Anti-NeuN antibody (Biotin) [EPR12763] (ab204681)

Properties

Applications

Our Abpromise guarantee covers the use of ab177487 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-FoFr 1/500 - 1/6000.
Flow Cyt 1/100.

ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-Fr 1/500.
IHC-P 1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified use at 1/800.

WB 1/1000 - 1/10000. Detects a band of approximately 48,50 kDa (predicted molecular weight: 34 kDa).

For unpurified use at 1/1000 - 1/2000.

ICC/IF 1/300.

For unpurified use at 1/80.

Target

Anti-NeuN antibody [EPR12763] - Neuronal Marker images

  • ab177487 staining NeuN in Mouse embryonic day 15 brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde, permeablized with 0.5% Triton X-100 in PBS, blocked with 10% serum for 1 hour at 25°C and antigen retrieval was by heat mediation in citrate buffer, pH 6. The sample was incubated with primary antibody (1/500 in PBS + 1% serum + 0.1% Triton X-100) for 16 hours at 25°C. An Alexa Fluor®594-conjugated Donkey anti-rabbit IgG (H+L) polyclonal (1/700) was used as the secondary antibody.

    See Abreview

  • ​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.

  • IHC image of NeuN (ab177487) with Anti-Rabbit IgG VHH Single Domain Antibody (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 1h 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, Anti-Rabbit IgG VHH Single Domain Antibody (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.

  • IHC-FoFr staining of NeuN staining on mouse pons sections using ab177487 (1/6000). The mouse was perfusion fixed using formaldehyde and 20µm sections were permeabilized with 0.5% tween. Blocking was performed using 1% BSA. ab177487 was diluted 1/6000 and incubated with the sections for 16 hours at 21°C. secondary antibody used was goat polyclonal to rabbit IgG conjugated to Alexa Fluor® 594 (1/1000).

    See Abreview

  • IHC-Fr staining of NeuN on zebrafish brain at 4dpf sections using ab177487 (1/100). The sections were fixed in paraformaldehyde and permeabilized using triton X. Antigen retrieval uisng sodium citrate was used. The sections were blocked using 5% BSA for 1 hour at 23°C. ab177487 was diluted 1/100 and incubated for 16 hours at 4°C. The secondary antibody used was anti rabbit IgG conjugated to Alexa Fluor® 488 (1/1000). Dapi used as counterstain.

    See Abreview

  • ​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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gliocytoma tissue labelling NeuN with unpurified ab177487 at 1/800. 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gliocytoma tissue labelling NeuN with purified ab177487 at 1/3000. 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 dog cerebellum 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 10mins 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).

  • 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 10mins 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 mouse brain (frontal cortex) sections using ab177487 (1/800). Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10mins 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).

  • 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 10mins 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).

  • 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 10mins 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 goat cerebellum 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 10mins 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).

  • IHC-P image of FOX3/NeuN staining on cat cerebellum sections using ab177487 (1/1000). Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10mins 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).

  • IHC-P image of FOX3/NeuN staining on sheep brain (Frontal cortex) sections using ab177487 (1/1000). Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10mins 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).

  • 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.

    See Abreview

  • 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 seurm 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.

    See Abreview

  • Immunocytochemsitry/Immunofluorescence analysis of SH-SY5Y cells labelling NeuN (green) with unpurified ab177487 at 1/80. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

  • Immunocytochemsitry/Immunofluorescence analysis of SH-SY5Y cells labelling NeuN (green) with purified ab177487 at 1/300. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

  • All lanes : Anti-NeuN antibody [EPR12763] - Neuronal Marker (ab177487) at 1/10000 dilution (purified)

    Lane 1 : Human fetal brain tissue lysate
    Lane 2 : 293 whole cell lysate
    Lane 3 : Mouse brain tissue lysate
    Lane 4 : Rat brain tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 34 kDa
    Observed band size : 46 kDa (why is the actual band size different from the predicted?)

    Exposure time -
    Lane 1-2: 3 minutes.
    Lane 3-4: 1 minute.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-NeuN antibody [EPR12763] - Neuronal Marker (ab177487) at 1 µg/ml (unpurified)

    Lane 1 : Brain (Mouse) Tissue Lysate
    Lane 2 : Cerebellum Mouse Tissue Lysate
    Lane 3 : Cerebellum Rat Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) at 1/10000 dilution

    Predicted band size : 34 kDa
    Observed band size : 48 + 50 kDa (why is the actual band size different from the predicted?)

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab177487 overnight at 4°C. Antibody binding was detected using ab175781 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

  • All lanes : Anti-NeuN antibody [EPR12763] - Neuronal Marker (ab177487) at 1/1500 dilution (unpurified)

    Lane 1 : Human fetal brain tissue lysate
    Lane 2 : Mouse brain tissue lysate
    Lane 3 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 34 kDa
    Observed band size : 46-48 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Overlay histogram showing U-87MG cells stained with ab177487 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. 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 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730, 1μg/1x106 cells used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • IHC-P image of NeuN (green) and GFAP (red) double staining on mouse cerebellum sections using ab177487 (1/5000) and 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 (ab4674) diluted at 1/1500. The primary antibody was detected using ab150097 Goat anti-rabbit IgG conjugated to Alexa Fluor® 488 (1/500) and ab150176 Goat anti-chicken IgY conjugated to Alexa Fluor® 594 (1/500)

References for Anti-NeuN antibody [EPR12763] - Neuronal Marker (ab177487)

This product has been referenced in:
  • Dekens DW  et al. Neutrophil Gelatinase-Associated Lipocalin and its Receptors in Alzheimer's Disease (AD) Brain Regions: Differential Findings in AD with and without Depression. J Alzheimers Dis 55:763-776 (2017). IHC-P ; Human . Read more (PubMed: 27716662) »
  • Srinageshwar B  et al. PAMAM Dendrimers Cross the Blood-Brain Barrier When Administered through the Carotid Artery in C57BL/6J Mice. Int J Mol Sci 18:N/A (2017). IHC-FoFr ; Mouse . Read more (PubMed: 28335421) »

See all 40 Publications for this product

Product Wall

Filter by Application

Filter by Species

Filter by Ratings

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH 6
Sample Mouse Tissue sections (Embryonic day 15 brain)
Specification Embryonic day 15 brain
Permeabilization Yes - 0.5% Triton X-100 in PBS
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted May 16 2014

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (Coronal Brain Sections)
Antigen retrieval step None
Permeabilization Yes - 0.3% Triton X in PBS
Specification Coronal Brain Sections
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Mr. Kyle Klingbeil

Verified customer

Submitted Aug 12 2015

Application Immunohistochemistry (Frozen sections)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: r.t°C
Sample Rat Tissue sections (Brain)
Specification Brain
Permeabilization Yes - 0.2% Triton X-100
Fixative Paraformaldehyde
Username

Dr. Fabio Canneva

Verified customer

Submitted Jul 09 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Serum as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 4°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer pH6.0
Sample Mouse Tissue sections (Mouse brain)
Specification Mouse brain
Permeabilization No
Fixative Paraformaldehyde
Username

Dr. Adam Walker

Verified customer

Submitted Jun 19 2014

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step Serum as blocking agent for 15 minute(s) · Concentration: 10% · Temperature: 22°C
Antigen retrieval step None
Sample Mouse Tissue sections (Brain)
Specification Brain
Permeabilization Yes - 0.5% Triton X
Fixative Paraformaldehyde
Username

Sumit Sarkar

Verified customer

Submitted May 21 2014

Application Immunohistochemistry (Frozen sections)
Blocking step Horse Seurm/ 0.4% Tx100 as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
Sample Mouse Tissue sections (Brain tissue)
Specification Brain tissue
Permeabilization No
Fixative Formaldehyde
Username

Ms. Eva Borger

Verified customer

Submitted May 15 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Ctric acid
Sample Cat Tissue sections (Cerebellum)
Specification Cerebellum
Permeabilization No
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Oct 29 2013

Application Immunocytochemistry/ Immunofluorescence
Sample Baboon Cell (Cortex)
Permeabilization No
Specification Cortex
Blocking step BSA as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: 24°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted May 05 2017

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
Username

Abcam user community

Verified customer

Submitted Mar 29 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Mouse Tissue sections (brain)
Antigen retrieval step None
Permeabilization Yes - 0.1% saponin
Specification brain
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Feb 23 2017

1-10 of 36 Abreviews

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"