Human ENO1 ELISA Kit, Fluorescent (ab229405)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 2.4 U/ml
- Range: 5.091 U/ml - 4200 U/ml
- Sample type: Cell culture extracts, Serum, Tissue Extracts
- Detection method: Fluorescent
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human ENO1 ELISA Kit, Fluorescent
See all ENO1 kits -
Detection method
Fluorescent -
Precision
Intra-assay Sample n Mean SD CV% HeLa extract 8 6.5% Inter-assay Sample n Mean SD CV% HeLa extract 3 11.4% -
Sample type
Serum, Cell culture extracts, Tissue Extracts -
Assay type
Sandwich (quantitative) -
Sensitivity
2.4 U/ml -
Range
5.091 U/ml - 4200 U/ml -
Recovery
Sample specific recovery Sample type Average % Range Serum 101 100% - 103% Cell culture extracts 93 92% - 93% Tissue Extracts 104 99% - 113% -
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.
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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.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 100X Stoplight Red Substrate 1 x 120µl 10X Human ENO1 Capture Antibody 1 x 600µl 10X Human ENO1 Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml Human ENO1 Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated Black 96-Well Microplate 1 unit Stoplight Red Substrate Buffer 1 x 12ml -
Research areas
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Function
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. 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. Stimulates immunoglobulin production.
MBP1 binds to the myc promoter and acts as a transcriptional repressor. May be a tumor suppressor. -
Tissue specificity
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. -
Pathway
Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 4/5. -
Sequence similarities
Belongs to the enolase family. -
Developmental stage
During 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. -
Post-translational
modificationsISGylated. -
Cellular localization
Nucleus and Cytoplasm. Cell membrane. Cytoplasm > myofibril > sarcomere > M line. Can translocate to the plasma membrane in either the homodimeric (alpha/alpha) or heterodimeric (alpha/gamma) form. ENO1 is localized to the M line. - Information by UniProt
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Alternative names
- 2 phospho D glycerate hydro lyase
- 2-phospho-D-glycerate hydro-lyase
- Alpha enolase
see all -
Database links
- Entrez Gene: 2023 Human
- Omim: 172430 Human
- SwissProt: P06733 Human
- Unigene: 517145 Human
Images
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Catchpoint ELISA Protocol Diagram
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.
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Example of human ENO1 standard curve in 1X Cell Extraction Buffer PTR.The ENO1 standard curve was prepared as described in Section 10. Raw data generated on SpectraMax M4 Multi-Mode Microplate Reader is shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native ENO1 in human HeLa cell extract based on a 50 µg/mL extract load and human liver tissue extract based on a 286 µg/mL extract load.The concentrations of ENO1 were measured in duplicate and interpolated from the ENO1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ENO1 concentration was determined to be 406 U/mL in HeLa cell extract and 441 U/mL in human liver tissue extract.
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Human recombinant ENO1 protein (600 ng/mL) and neuron-specific human recombinant ENO2 protein (600 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 the background. Note that this kit does not detect ENO2 protein.
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Extracts of human liver and human heart were analyzed with this kit.The concentrations of ENO1 were interpolated from the ENO1 standard curve and graphed in U of ENO1 per mg of extract (mean +/- SD, n=2). Since striated muscles of myocardium contain ENO3 homodimers and ENO1/ENO3 heterodimers, the data suggest that the kit is not reactive with muscle specific ENO3.
