< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
No significant interference observed with available related molecules.
QC83 , Quantikine Immunoassay Control Set 726 for Human Enolase 2 -
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
The recovery of Enolase 2 spiked to levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=8)
Cell Lysates (n=8)
Platelet-poor Heparin Plasma (n=4)
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.
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.
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.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
100 µL Assay Diluent
Add 100 µL of Assay Diluent to each well.
50 µL Standard, Control, or Sample
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.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
200 µL Conjugate
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.
Aspirate and wash 4 times.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
50 µL Stop Solution
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.
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