Overview

  • Product name
    Human ENO1 ELISA Kit (Alpha-Enolase)
  • Detection method
    Colorimetric
  • 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
    8.1 U/ml
  • Range
    12.5 U/ml - 800 U/ml
  • Recovery

    90.1 %

    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

    Abcam’s ENO1 (alpha-enolase) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of ENO1 protein in Human serum, 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.

  • Notes

    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
    Microplate

Properties

Applications

Our Abpromise guarantee covers the use of ab181417 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

  • ELISA Protocol Summary
  • Background-subtracted data values (mean +/- SD, n=2) are graphed.

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

  • Assay specificity. 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.

  • Assay specificity. 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

ab181417 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Answer

I can confirm both sample types (serum or cell and tissue extracts) and standard should be diluted in 1X Extraction Buffer PTR. This is how the lab validated the assay.

Diluent NS is supplied  in all our SimpleStep kits although there is no use of the Sample Diluent NS in this particular assay.

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