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Human NSE(Neuron Specific Enolase) ELISA Kit

Citations(7)Uniprot : P09104

Cat.No.:E-EL-H1047
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	2.34-150 ng/mL
Sensitivity	1.40 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human NSE in samples. No significant cross-reactivity or interference between Human NSE and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human NSE concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human NSE. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human NSE and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human NSE, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human NSE. You can calculate the concentration of Human NSE in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human NSE were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human NSE were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	7.40	23.30	55.20	7.60	23.70	59.30
Standard deviation	0.40	1.10	2.20	0.50	1.20	2.70
CV (%)	5.41	4.72	3.99	6.58	5.06	4.55

Recovery

The recovery of Human NSE spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	94-108	102
EDTA plasma (n=8)	84-99	90
Cell culture media (n=8)	88-100	95

Linearity

Samples were spiked with high concentrations of Human NSE and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P09104
Synonyms	Enolase γ, Enolase 2
Research Area	Cancer, Metabolism, Developmental biology, Neuroscience, Stem cells, Tags and Cell Markers

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

JOURNAL OF MEDICAL VIROLOGY (2022) IF: 20.693
Neurological symptoms and neuronal damage markers in acute COVID-19: Is there a correlation? A pilot study

DOI: 10.1002/jmv.28240

PMID: 36262025
Sample: serum
INTERNATIONAL JOURNAL OF DEVELOPMENTAL NEUROSCIENCE (2017) IF: 2.046
Evaluation of serum Neuron-specific enolase, S100B, myelin basic protein and glial fibrilliary acidic protein as brain specific proteins in children with autism spectrum disorder

DOI: 10.1016/j.ijdevneu.2017.06.011

Sample: Serum
BRAIN INJURY (2021) IF: 2.311
Changes in amplitude-integrated electroencephalography, neuron-specific enolase, and S100B in neonates with brain injury induced by neonatal hyperbilirubinemia and their significance

DOI: 10.1080/02699052.2021.1931449

PMID: 34097553
Sample: serum
BMC Neurology (2022) IF: 2.474
Changes of biochemical biomarkers in the serum of children with convulsion status epilepticus: a prospective study

DOI: 10.1186/s12883-022-02686-2

PMID: 35624413
Sample: serum
INTERNATIONAL JOURNAL OF ENVIRONMENTAL ANALYTICAL CHEMISTRY (2019) IF: 1.267
An ultrasensitive immunosensor based on tri-armed star poly(glycidyl methacrylate) polymer-coated ITO-PET electrode for detection of neuron-specific enolase in human serum

DOI: 10.1080/03067319.2019.1673744

Sample: serum
MEDICINE (2021) IF: 1.889
Serum NSE and S100B protein levels for evaluating the impaired consciousness in patients with acute carbon monoxide poisoning

DOI: 10.1097/MD.0000000000026458

PMID: 34160445
Sample: serum
BMC Research Notes (2022)
Plasma Neuron-Specific Enolase is not a reliable biomarker for staging Trypanosoma brucei rhodesiense sleeping sickness patients

DOI: 10.1186/s13104-022-05981-w

Sample: Plasma

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