Overview

  • Product name

    Human ENO1 ELISA Kit, Fluorescent
    See all ENO1 kits
  • Detection method

    Fluorescent
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    HeLa extract 5 5.7%
    Inter-assay
    Sample n Mean SD CV%
    HeLa extract 3 4.7%
  • Sample type

    Serum, Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    2.4 U/ml
  • Range

    3.13 U/ml - 800 U/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 96.4 87.4% - 104.1%
    Cell culture media 99.5 97.2% - 102.7%
    Goat Serum 90.4 83.5% - 95.2%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    ENO1 (Alpha-Enolase) in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of ENO1 (Alpha-Enolase) protein in humanserum, cell and tissue extracts.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.


    The CatchPoint 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. CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.

  • Notes

    Enolase is a multifunctional enzyme that, as well as its role in glycolysis, plays a part in various processes such as growth control, hypoxia tolerance and allergic responses. It may also function in the intravascular and pericellular fibrinolytic system due to its ability to serve as a receptor and activator of plasminogen on the cell surface of several cell-types such as leukocytes and neurons. It stimulates immunoglobulin production. Mammalian enolase is composed of 3 isozyme subunits, alpha (ENO1), beta (ENO3) and gamma (ENO2), which can form homodimers or heterodimers which are cell-type and development-specific. The 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.

    Alpha enolase (ENO1, P06733) ENO1 is localized to cytoplasm. It can translocate to the plasma membrane in either the homodimeric (alpha/alpha) or heterodimeric (alpha/gamma) form. MBP1, a shorter isoform of the ENO1, binds to the myc promoter and acts as a transcriptional repressor. It may be a tumor suppressor. It locates mainly in nucleus. ENO1 is used as a diagnostic marker for many tumors and, in the heterodimeric form, alpha/gamma, as a marker for hypoxic brain injury after cardiac arrest. Also it is a marker for endometriosis. Antibodies against ENO1 are present in sera from patients with cancer-associated retinopathy syndrome (CAR), a progressive blinding disease which occurs in the presence of systemic tumor growth, primarily small-cell carcinoma of the lung and other malignancies. ENO1 is identified as an autoantigen in Hashimoto encephalopathy (HE) a rare autoimmune disease associated with Hashimoto thyroiditis (HT). HT is a disorder in which destructive processes overcome the potential capacity of thyroid replacement leading to hypothyroidism.

  • 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 ab229405 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

  • 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.

     

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

  • Background subtracted data values (mean +/- SD, n=2) are plotted.

  • Human recombinant ENO1 protein (800 ng/mL) and the neuron-specific human recombinant ENO2 protein (800 ng/mL) were analyzed with this kit. Background subtracted data values (mean +/- SD, n=2) are plotted. The dotted line represents O.D. (450 nm) value corresponding to MDD. Note that this kit does not detect ENO2 protein.

  • 3 µg/mL of extracts of human liver homogenate (HLH) and human heart homogenate (HHH) were analyzed with this kit. The concentrations of ENO1 were interpolated from data values using ENO1 standard curve and graphed in U of ENO1 per µg of extract (mean +/- SD, n=2). The dotted line represents MDD of the assay. The data shows that ENO1 is not detectable in HHH. Since striated muscles of myocardium contain ENO3 homodimers and ENO1/ENO3 heterodimers, the data strongly suggest that the kit is not reactive with muscle-specific ENO3.

Protocols

References

ab229405 has not yet been referenced specifically in any publications.

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