Overview

  • Product name
    Anti-NeuN antibody [EPR12763] - BSA and Azide free
    See all NeuN primary antibodies
  • Description
    Rabbit monoclonal [EPR12763] to NeuN - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-FoFr, Flow Cyt, IHC-Fr, IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Goat, Cat, Dog, Human, Zebrafish, Common marmoset
  • Immunogen

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

  • Positive control
    • This antibody gave a positive signal in the following tissue lysates: Mouse Brain; Mouse Cerebellum; Rat cerebellum; Human Fetal brain as well as the following whole cell lysates: 293T, HeLa and SH-SY5Y lysates; SH-SY-5Y cells. Flow Cyt: U-87MG cells.
  • General notes

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab209898 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 Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-Fr Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

WB Use at an assay dependent concentration. Detects a band of approximately 48.50 kDa (predicted molecular weight: 34 kDa).
ICC/IF Use at an assay dependent concentration.

Target

Images

  • 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 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730, 1μg/1x106 cells used under the same conditions. Unlabeled 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • Immunocytochemsitry/Immunofluorescence analysis of SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • Immunocytochemsitry/Immunofluorescence analysis of SH-SY5Y (Human neuroblastoma cell line from bone marrow) 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • ?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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • IHC-Fr staining of NeuN on zebrafish brain tissue at 4 days post-fertilization 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • ?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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • 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)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • 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 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • 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 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • 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 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • 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 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

  • 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 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab177487).

References

This product has been referenced in:
  • Borger E  et al. The calcium-binding protein EFhd2 modulates synapse formation in vitro and is linked to human dementia. J Neuropathol Exp Neurol 73:1166-82 (2014). IHC . Read more (PubMed: 25383639) »
  • Gondo A  et al. Sustained Down-regulation of ß-Dystroglycan and Associated Dysfunctions of Astrocytic Endfeet in Epileptic Cerebral Cortex. J Biol Chem 289:30279-88 (2014). IHC-P ; Mouse . Read more (PubMed: 25228692) »
See all 2 Publications for this product

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