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Mouse NOS3/eNOS(Nitric Oxide Synthase 3, Endothelial) ELISA Kit

Citations(6)Uniprot : P70313

Cat.No.:E-EL-M0456
Reactivity: Mouse

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	15.63-1000 pg/mL
Sensitivity	9.38 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Mouse NOS3/eNOS in samples. No significant cross-reactivity or interference between Mouse NOS3/eNOS 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 Mouse NOS3/eNOS 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 Mouse NOS3/eNOS. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse NOS3/eNOS 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 Mouse NOS3/eNOS, 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 Mouse NOS3/eNOS. You can calculate the concentration of Mouse NOS3/eNOS 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 Mouse NOS3/eNOS were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse NOS3/eNOS 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 (pg/mL)	45.50	109.20	426.30	44.30	119.40	394.80
Standard deviation	3.20	5.10	17.90	2.40	6.20	18.60
CV (%)	7.03	4.67	4.20	5.42	5.19	4.71

Recovery

The recovery of Mouse NOS3/eNOS 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)	93-108	99
EDTA plasma (n=8)	90-104	97
Cell culture media (n=8)	92-104	99

Linearity

Samples were spiked with high concentrations of Mouse NOS3/eNOS and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P70313
Synonyms	ECNOS, NOS III, E-NOS
Research Area	Cancer, Cell Biology, Cardiovascular, Metabolism, Neuroscience

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

Frontiers in Cardiovascular Medicine (2021) IF: 6.05
Potential Mechanism of Dingji Fumai Decoction Against Atrial Fibrillation Based on Network Pharmacology, Molecular Docking, and Experimental Verification Integration Strategy

DOI: 10.3389/fcvm.2021.712398

PMID: 34859062
Sample: HL-1 cells
Stem Cell Research & Therapy (2021) IF: 6.832
Sox9-expressing cells promote regeneration after radiation-induced lung injury via the PI3K/AKT pathway

DOI: 10.1186/s13287-021-02465-9

PMID: 34215344
Sample: serum
Frontiers in Pharmacology (2021) IF: 5.811
Ginsenoside Rg1-Notoginsenoside R1-Protocatechuic Aldehyde Reduces Atherosclerosis and Attenuates Low-Shear Stress-Induced Vascular Endothelial Cell Dysfunction

DOI: 10.3389/fphar.2020.588259

PMID: 33568993
Sample: Serum
TOXICOLOGY IN VITRO (2021) IF: 3.5
Triclosan affects the expression of nitric oxide synthases (NOSs), peroxisome proliferator-activated receptor gamma (PPARγ), and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in mouse neocortical neurons in vitro

DOI: 10.1016/j.tiv.2021.105143

PMID: 33722737
Sample: Cell culture supernatant
Evidence-based Complementary and Alternative Medicine (2022) IF: 2.63
Biejiajian Pill Ameliorates Diabetes-Associated Atherosclerosis through Inhibition of the NLRP3 Inflammasome

DOI: 10.1155/2022/9131178

PMID: 35692570
Sample: serum
NEUROCHEMICAL RESEARCH (2019) IF: 2.782
The VGVAPG Peptide Regulates the Production of Nitric Oxide Synthases and Reactive Oxygen Species in Mouse Astrocyte Cells In Vitro

DOI: 10.1007/s11064-019-02746-z

PMID: 30759294
Sample: Cell lysate

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