SimpleStep

Human Neuron specific Enolase ELISA Kit (ab217778)

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Overview

  • Product name

    Human Neuron specific Enolase ELISA Kit
    See all NSE kits

  • Detection method

    Colorimetric

  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 5 4.3%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 11.4%

  • Sample type

    Cell culture supernatant, Serum, Cell culture extracts, Tissue Extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma

  • Assay type

    Sandwich (quantitative)

  • Sensitivity

    23 pg/ml

  • Range

    312.5 pg/ml - 20000 pg/ml

  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 106 103% - 108%
    Cell culture extracts 90 89% - 91%
    Tissue Extracts 110 106% - 113%
    Cell culture media 103 102% - 105%
    Heparin Plasma 109 104% - 114%
    EDTA Plasma 105 96% - 118%
    Citrate Plasma 102 100% - 105%

  • Assay time

    1h 30m

  • Assay duration

    One step assay

  • Species reactivity

    Reacts with: Mouse, Human

  • Product overview

    Neuron-specific Enolase in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Neuron-specific Enolase protein in serum, plasma, cell culture supernatant, cell and tissue extracts. 


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Sensitivity:


    Samples in 1X Cell Extraction Buffer PTR: 23 pg/mL


    Samples in Sample Diluent NS: 72 pg/mL

  • Notes

    Neuron-specific Enolase (also known as NSE, gamma-enolase and Enolase 2) is a cytoplasmic phosphopyruvate hydratase.  Neuron-specific Enolase has two related family members, Enolase 1 and Enolase 3.  Neuron-specific Enolase can be used to identify neuronal cells and normal or malignant cells with neuroendocrine origin.

  • Tested applications

    Suitable for: Sandwich ELISAmore details

  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

Applications

Our Abpromise guarantee covers the use of ab217778 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • Other - Human Neuron specific Enolase ELISA Kit (ab217778)
    Other - Human Neuron specific Enolase ELISA Kit (ab217778)

    SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Human Neuron specific Enolase standard curve comparison data
    Human Neuron specific Enolase standard curve comparison data

    Standard curve comparison between human Neuron specific Enolase SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows comparable sensitivity.

  • Example of human Neuron-specific Enolase standard curve in Sample Diluent NS.
    Example of human Neuron-specific Enolase standard curve in Sample Diluent NS.

    Background-subtracted data values (mean +/- SD) are graphed.

  • Example of human Neuron-specific Enolase standard curve in 1X Cell Extraction Buffer PTR.
    Example of human Neuron-specific Enolase standard curve in 1X Cell Extraction Buffer PTR.

    Background-subtracted data values (mean +/- SD) are graphed.

  • Interpolated concentrations of spike Neuron-specific Enolase in human serum, and plasma samples.
    Interpolated concentrations of spike Neuron-specific Enolase in human serum, and plasma samples.

    The concentrations of Neuron-specific Enolase were measured in duplicates, interpolated from the Neuron-specific Enolase standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 25%, plasma (EDTA) 25%, plasma (heparin) 25%, and plasma (citrate) 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

  • Interpolated concentrations of native Neuron-specific Enolase in human serum, and plasma samples.
    Interpolated concentrations of native Neuron-specific Enolase in human serum, and plasma samples.

    The concentrations of Neuron-specific Enolase were measured in duplicates, interpolated from the Neuron-specific Enolase standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 25%, plasma (EDTA) 25%, plasma (heparin) 25%, and plasma (citrate) 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Neuron-specific Enolase concentration was determined to be 821 pg/mL in serum, 8545 pg/mL in plasma (EDTA), 1194 pg/mL in plasma (heparin), and 3386 pg/mL in plasma (citrate).

  • Serum from ten individual healthy human female donors was measured in duplicate.
    Serum from ten individual healthy human female donors was measured in duplicate.

    Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Eight out of ten donors tested below the detectable dose.  The mean Neuron-specific Enolase concentration of two donors was determined to be 1553 pg/mL with a range of 766 – 2340 pg/mL.

  • Interpolated concentrations of native Neuron-specific Enolase in human peripheral blood mononuclear (PBMC) cell culture supernatant samples.
    Interpolated concentrations of native Neuron-specific Enolase in human peripheral blood mononuclear (PBMC) cell culture supernatant samples.

    PBMCs were grown in the absence (unstimulated) or presence (stimulated) of phytohemagllutinin (PHA) for 3 days.  The concentrations of Neuron-specific Enolase were measured in duplicates, interpolated from the Neuron-specific Enolase standard curves and corrected for sample dilution. Undiluted samples are as follows: unstimulated 50%, and stimulated 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Neuron-specific Enolase concentration was determined to be undetectable in media, 711 pg/mL in unstimulated, and 4051 pg/mL in stimulated.

  • Interpolated concentrations of native Neuron-specific Enolase in human SH-SY5Y cell extract and human brain homogenate tissue extract samples based on a 50 µg/mL and 100 µg/mL extract load.
    Interpolated concentrations of native Neuron-specific Enolase in human SH-SY5Y cell extract and human brain homogenate tissue extract samples based on a 50 µg/mL and 100 µg/mL extract load.

    The concentrations of Neuron-specific Enolase were measured in duplicate and interpolated from the Neuron-specific Enolase standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Neuron-specific Enolase concentration was determined to be 5828 pg/mL in SH-SY5Y cell extract and 1482 pg/mL in human brain homogenate tissue extract.

Protocols

References

This product has been referenced in:

  • Zhu J  et al. Effects of neuritin on the differentiation of bone marrow-derived mesenchymal stem cells into neuron-like cells. Mol Med Rep 16:3201-3207 (2017). Read more (PubMed: 28714031) »
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