Human Enolase 2/Neuron-specific Enolase Quantikine ELISA Kit

(1 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Serum (50 uL), Platelet-poor Heparin Plasma (50 uL)
  • Sensitivity
    0.038 ng/mL
  • Assay Range
    0.3 - 20 ng/mL (Cell Culture Supernates, Cell Lysates, Serum, Platelet-poor Heparin Plasma)
  • Specificity
    Natural and recombinant human Enolase 2
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC83, Quantikine Immunoassay Control Set 726 for Human Enolase 2 - Please Inquire
Product Summary
The Quantikine Human Enolase 2 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Enolase 2 in cell culture supernates, cell lysates, serum, and plasma. It contains E. coli-expressed recombinant human Enolase 2 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Enolase 2 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human Enolase 2.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Cell Lysates, Serum, Platelet-poor Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean2.158.41131.968.512.6
Standard Deviation0.030.170.360.130.330.55
CV%1.422.86.73.94.3

Recovery

The recovery of Enolase 2 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=8) 99 85-114
Cell Lysates (n=8) 105 85-115
Platelet-poor Heparin Plasma (n=4) 95 85-111
Serum (n=4) 95 85-110
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Enolase 2 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Enolase 2/Neuron-specific Enolase Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Enolase 2/Neuron-specific Enolase

Enolase 2 (2-phospho-D-glycerate hydrolyase; also Neuron-specific Enolase, NSE, neural enolase and gamma-enolase) is a 46 kDa member of the Enolase family of enzymes. It is expressed in developing neurons and glia, is known to catalyze the generation of phosphoenolpyruvate, and is suggested to possess neurotrophic activity for neurons, likely through an extracellular mechanism. Human Enolase 2 is 434 amino acids (aa) in length. The enzymatic site spans most of the length of the molecule. Enolase 2 exists as both a noncovalently-linked homodimer, or heterodimer with alpha-enolase. Full-length human Enolase 2 shares 99% aa identity with both mouse and canine Enolase 2. It shares 83% aa identity with human enolases 1 and 3.

    • Entrez Gene IDs
      2026 (Human); 13807 (Mouse); 24334 (Rat);
    • Alternate Names
      2-phospho-D-glycerate hydrolyase; 2-phospho-D-glycerate hydro-lyase; EC 4.2.1.11; ENO2; enolase 2 (gamma, neuronal); Enolase 2; gamma-Enolase; Neural enolase; neuron specific gamma enolase; neurone-specific enolase; Neuronspecific Enolase; Neuron-specific enolase; NSE;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 100 µL Assay Diluent
    4.   Add 100 µL of Assay Diluent to each well.

    5. 50 µL Standard, Control, or Sample
    6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
    10.   Aspirate and wash 4 times.

    11. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    Citations:

    R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

    1 Citations: Showing 1 - 1

    1. Cerebral Biomarkers in Women With Preeclampsia Are Still Elevated 1 Year Postpartum
      Am J Hypertens, 2016;0(0):.
      Species: Human
      Sample Type: Plasma
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 726 for Human Enolase 2

    Ctrl QC83
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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    Cell Lysis Buffer 1 (1 x 21 mL)

    ELISA 890713 1
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