Overview

  • Product nameAnti-NSE antibody [85F11]
    See all NSE primary antibodies
  • Description
    Mouse monoclonal [85F11] to NSE
  • SpecificityWe have data to indicate that this antibody may not cross react with Rat. However, this has not been conclusively tested and expression levels may vary in certain cell lines/tissues.
  • Tested applicationsSuitable for: Flow Cyt, IHC-FoFr, ICC/IF, ELISA, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Goat, Chicken, Macaque Monkey
  • Immunogen

    Recombinant full length protein purified from E.coli (Human).

  • Positive control
    • SH-SY5Y whole cell lysate. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: SKNSH.

Properties

Applications

Our Abpromise guarantee covers the use of ab16808 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
Flow Cyt Use 1µg for 106 cells. (paraformaldehyde fixed cells)



ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
IHC-FoFr 1/100. PubMed: 19279043
ICC/IF Use a concentration of 10 µg/ml.
ELISA Use at an assay dependent concentration.
WB 1/2000. Predicted molecular weight: 48 kDa.

Target

  • FunctionHas neurotrophic and neuroprotective properties on a broad spectrum of central nervous system (CNS) neurons. Binds, in a calcium-dependent manner, to cultured neocortical neurons and promotes cell survival.
  • Tissue specificityThe alpha/alpha homodimer is expressed in embryo and in most adult tissues. The alpha/beta heterodimer and the beta/beta homodimer are found in striated muscle, and the alpha/gamma heterodimer and the gamma/gamma homodimer in neurons.
  • PathwayCarbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 4/5.
  • Sequence similaritiesBelongs to the enolase family.
  • Developmental stageDuring ontogenesis, there is a transition from the alpha/alpha homodimer to the alpha/beta heterodimer in striated muscle cells, and to the alpha/gamma heterodimer in nerve cells.
  • Cellular localizationCytoplasm. Cell membrane. Can translocate to the plasma membrane in either the homodimeric (alpha/alpha) or heterodimeric (alpha/gamma) form.
  • Information by UniProt
  • Database links
  • Alternative names
    • 2 phospho D glycerate hydrolyase antibody
    • 2-phospho-D-glycerate hydro-lyase antibody
    • Eno 2 antibody
    • ENO2 antibody
    • ENOG antibody
    • ENOG_HUMAN antibody
    • Enolase 2 (gamma, neuronal) antibody
    • Enolase 2 antibody
    • Enolase 2 gamma neuronal antibody
    • Enolase2 antibody
    • Epididymis secretory protein Li 279 antibody
    • Gamma enolase antibody
    • Gamma-enolase antibody
    • HEL S 279 antibody
    • Neural enolase antibody
    • Neuron specific enolase antibody
    • Neuron specific gamma enolase antibody
    • Neuron-specific enolase antibody
    • Neurone specific enolase antibody
    • NSE antibody
    see all

Anti-NSE antibody [85F11] images

  • ICC/IF image of ab16808 stained SKNSH cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab16808 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.



  • Predicted band size : 48 kDa
    ab16808 at a 1/2000 dilution staining approx. 48kDa NSE in 1) SH-SY5Y and 2) U87MG cell lysates by Western blot (ECL).  ab16808 at a 1/2000 dilution staining approx. 48kDa NSE in 1) SH-SY5Y and 2) U87MG cell lysates by Western blot (ECL).
  • Overlay histogram showing SH-SY5Y cells stained with ab16808 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16808, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in SH-SY5Y cells fixed with methanol (5 min) used under the same conditions.

    Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.

References for Anti-NSE antibody [85F11] (ab16808)

This product has been referenced in:
  • Fatemi SH  et al. Expression of GABAA a2-, ß1- and ?-receptors are altered significantly in the lateral cerebellum of subjects with schizophrenia, major depression and bipolar disorder. Transl Psychiatry 3:e303 (2013). WB ; Human . Read more (PubMed: 24022508) »
  • Yu Y  et al. Evaluation of blastomere biopsy using mouse model indicates the potential high-risk of neurodegenerative disorders in the offspring. Mol Cell Proteomics : (2009). WB, IHC-FoFr ; Mouse . Read more (PubMed: 19279043) »

See all 3 Publications for this product

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Thank you for contacting Abcam and providing your protocol information.

Using mouse primary antibodies against mouse cells for flow cytometry is not ideal, as the anti-mouse secondary is susceptible to binding to the mouse cells non-specifical...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (frontal cortex)
Loading amount 30 µg
Specification frontal cortex
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Oct 07 2009

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