Human ENO1 ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human ENO1 with a sensitivity of 2.4 U/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Application | Reactivity | Dilution info | Notes |
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Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Glycolytic enzyme the catalyzes the conversion of 2-phosphoglycerate to phosphoenolpyruvate (PubMed:1369209, PubMed:29775581). In addition to glycolysis, involved in various processes such as growth control, hypoxia tolerance and allergic responses (PubMed:10802057, PubMed:12666133, PubMed:2005901, PubMed:29775581). 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 (PubMed:12666133). Stimulates immunoglobulin production (PubMed:1369209). Isoform MBP-1. Binds to the myc promoter and acts as a transcriptional repressor. May be a tumor suppressor.
ENO1L1, MBPB1, MPB1, ENO1, Alpha-enolase, 2-phospho-D-glycerate hydro-lyase, C-myc promoter-binding protein, Enolase 1, MBP-1, MPB-1, Non-neural enolase, Phosphopyruvate hydratase, Plasminogen-binding protein, NNE
Human ENO1 ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human ENO1 with a sensitivity of 2.4 U/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Sample | n | C.V. |
---|---|---|
Sample HeLa Extract | n 8 | C.V. 6.5 |
Sample | n | C.V. |
---|---|---|
Sample HeLa Extract | n 3 | C.V. 11.4 |
Sample type | Average % | Range |
---|---|---|
Sample type Serum | Average % = 101 | Range 100 - 103 % |
Sample type Cell culture extracts | Average % = 93 | Range 92 - 93 % |
Sample type Tissue Extracts | Average % = 104 | Range 99 - 113 % |
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.
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.
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Enolase 1 or alpha-enolase is an important enzyme in the glycolytic pathway. It catalyzes the conversion of 2-phosphoglycerate to phosphoenolpyruvate. We also refer to it as an enolase reaction. ENO1 protein has a molecular mass of approximately 47 kDa and is expressed in most tissues but found most abundantly in the brain muscle and liver. Additionally ENO1 can exist in both cytosolic and plasma membrane-associated forms playing functional roles in different cellular contexts.
The ENO1 protein serves multiple functions beyond its glycolytic activity. It also acts as a plasminogen receptor on the cell surface aiding in the modulation of pericellular and extracellular matrix degradation processes. Through this role ENO1 influences cell migration and tissue remodeling. This protein can form part of larger protein complexes contributing to its versatility in various cellular functions.
ENO1 plays a critical role in glycolysis a central metabolic pathway that converts glucose into pyruvate generating ATP as a result. In this pathway ENO1 closely interacts with other enzymes like phosphoglycerate kinase and pyruvate kinase. Additionally as a plasminogen receptor ENO1 links to the plasminogen activation pathway affecting fibrinolysis and cellular response to tissue injury.
ENO1 is associated with cancer and neurodegenerative diseases. Overexpression or aberrant regulation of ENO1 often correlates with tumor progression and metastasis in cancers such as lung and breast cancer. In neurodegenerative disorders like Alzheimer's disease abnormal functioning of ENO1 can impact brain energy metabolism and neuron degeneration. ENO1's interaction with other proteins such as beta-enolase further connects it to these conditions influencing disease mechanisms and patient outcomes.
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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.
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.
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.
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.
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