Human Enolase 2/Neuron-specific Enolase Quantikine ELISA Kit

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Human Enolase 2/Neuron-specific Enolase ELISA Standard Curve
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Human Enolase 2/Neuron-specific Enolase Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
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)
0.038 ng/mL
Assay Range
0.3 - 20 ng/mL (Cell Culture Supernates, Cell Lysates, Serum, Platelet-poor Heparin Plasma)
Natural and recombinant human Enolase 2
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
No significant interference observed with available related molecules.

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.


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
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 2.15 8.41 13 1.96 8.5 12.6
Standard Deviation 0.03 0.17 0.36 0.13 0.33 0.55
CV% 1.4 2 2.8 6.7 3.9 4.3


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


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 ELISA Linearity

Scientific Data

Human Enolase 2/Neuron-specific Enolase ELISA Standard Curve

Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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; ENO2; enolase 2 (gamma, neuronal); Enolase 2; gamma-Enolase; Neural enolase; neuron specific gamma enolase; neurone-specific enolase; Neuronspecific Enolase; Neuron-specific enolase; NSE
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to

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 for Human Enolase 2/Neuron-specific Enolase Quantikine ELISA Kit

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.

4 Citations: Showing 1 - 4
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  1. Blood-brain barrier and gut barrier dysfunction in chronic kidney disease with a focus on circulating biomarkers and tight junction proteins
    Authors: L Hernandez, LJ Ward, S Arefin, T Ebert, A Laucyte-Ci, GOING-FWD, O Heimbürger, P Barany, L Wennberg, P Stenvinkel, K Kublickien
    Scientific Reports, 2022;12(1):4414.
    Species: Human
    Sample Types: Serum
  2. New Insights on Old Biomarkers Involved in Tumor Microenvironment Changes and Their Diagnostic Relevance in Non-Small Cell Lung Carcinoma
    Authors: K Wadowska, P B?asiak, A Rzechonek, I Bil-Lula, M ?liwi?ska-
    Biomolecules, 2021;11(8):.
    Species: Human
    Sample Types: Serum
  3. Effects of Kudiezi Injection on Serum Inflammatory Biomarkers in Patients with Acute Cerebral Infarction
    Authors: X Liu, X Jin, B Chen, X Liu, X Liang, X Fang, H Wu, X Fu, H Zheng, X Ding, N Duan, Y Zhang
    Dis. Markers, 2018;2018(0):7936736.
    Species: Human
    Sample Types: Serum
  4. Cerebral Biomarkers in Women With Preeclampsia Are Still Elevated 1 Year Postpartum
    Am J Hypertens, 2016;0(0):.
    Species: Human
    Sample Types: Plasma


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