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Human NE/ELA2(Neutrophil Elastase/Elastase-2) ELISA Kit

Citations(7)Uniprot : P08246

Cat.No.:E-EL-H1946
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.78-50 ng/mL
Sensitivity	0.47 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human NE/ELA2 in samples. No significant cross-reactivity or interference between Human NE/ELA2 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 NE/ELA2 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 NE/ELA2. 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 NE/ELA2 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 NE/ELA2, 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 NE/ELA2. You can calculate the concentration of Human NE/ELA2 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 NE/ELA2 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human NE/ELA2 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)	2.58	5.84	23.39	2.53	6.40	25.58
Standard deviation	0.13	0.31	0.91	0.17	0.28	1.21
CV (%)	5.04	5.31	3.89	6.72	4.38	4.73

Recovery

The recovery of Human NE/ELA2 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)	89-103	94
EDTA plasma (n=8)	88-100	94
Cell culture media (n=8)	93-109	100

Linearity

Samples were spiked with high concentrations of Human NE/ELA2 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P08246
Research Area	Cancer, Immunology, Microbiology, Signal transduction

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 CELLULAR AND MOLECULAR MEDICINE (2021) IF: 5.31
Identification of key candidate biomarkers for severe influenza infection by integrated bioinformatical analysis and initial clinical validation

DOI: 10.1111/jcmm.16275

PMID: 33448094
Sample: Plasma
ACTA DIABETOLOGICA (2022) IF: 4.28
Deep transcranial magnetic stimulation in combination with skin thermography in obesity: a window on sympathetic nervous system

DOI: 10.1007/s00592-022-01859-2

PMID: 35174415
Sample: blood
JOURNAL OF VASCULAR SURGERY (2020) IF: 3.405
Magnetic resonance imaging assessment of proteolytic enzyme concentrations and biologic properties of intraluminal thrombus in abdominal aortic aneurysms

DOI: 10.1016/j.jvs.2019.11.032

Sample: aortic wall
JOURNAL OF INTERFERON AND CYTOKINE RESEARCH (2022) IF: 3.657
Can the Prognosis of COVID-19 Disease Be Determined by Fecal Markers and Cytokines?

DOI: 10.1089/jir.2022.0098

PMID: 36179036
Sample: fecal
Immunity Inflammation and Disease (2021) IF: 2.239
Neutrophil extracellular traps induce thrombogenicity in severe carotid stenosis

DOI: 10.1002/iid3.466

PMID: 34102007
Sample: plasma
ANNALS OF VASCULAR SURGERY (2022) IF: 1.607
Correlation between proteolytic activity and abdominal aortic aneurysm wall morphology with intraluminal thrombus volume

DOI: 10.1016/j.avsg.2022.05.039

PMID: 35779804
Sample: Abdominal Aortic Aneurysm (AAA) Wall
MEDICINE (2016) IF: 1.889
Disequilibrium of Blood Coagulation and Fibrinolytic System in Patients With Coronary Artery Ectasia

DOI: 10.1097/MD.0000000000002779

Sample: plasma

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