ELISA Kit for Endothelial NOS (eNOS) Rattus norvegicus (Rat) Sandwich ELISA

NOS3; ECNOS; NOS III; E-NOS; C-NOS; ENOS; CNOS; Constitutive NOS; Nitric Oxide Synthase 3, Endothelial

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  • ELISA Kit for Endothelial NOS (eNOS) Packages (Simulation)
  • ELISA Kit for Endothelial NOS (eNOS) Packages (Simulation)
  • ELISA Kit for Endothelial NOS (eNOS) Results demonstration
  • SEA868Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Endothelial NOS (eNOS) and the recovery rates were calculated by comparing the measured value to the expected amount of Endothelial NOS (eNOS) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 96-105 101
EDTA plasma(n=5) 88-97 92
heparin plasma(n=5) 86-101 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Endothelial NOS (eNOS) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Endothelial NOS (eNOS) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Endothelial NOS (eNOS) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 81-95% 78-96% 80-98% 80-94%
EDTA plasma(n=5) 88-97% 84-99% 89-101% 80-103%
heparin plasma(n=5) 91-99% 84-94% 87-96% 91-103%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Endothelial NOS (eNOS)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Endothelial NOS (eNOS). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Endothelial NOS (eNOS). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Endothelial NOS (eNOS), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Endothelial NOS (eNOS) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Impact of matrine on inflammation related factors in rat intestinal microvascular endothelial cellsScienceDirect: S0378874109004541
  • A Novel Mechanism of Formaldehyde Neurotoxicity: Inhibition of Hydrogen Sulfide Generation by Promoting Overproduction of Nitric OxidePubMed: 23359814
  • Inflammation influence on the formation of endothelial dysfunction in hypertensive patients caused by obesityKnmu: Source
  • SIRT1 Gene Polymorphisms Affect the Protein Expression in Cardiovascular DiseasesPlos: Source
  • L-ARGININE, BUT NOT L-NAME PROTECTS AGAINST LIVER INJURY INDUCED BY EXPERIMENTAL ISCHEMIA-REPERFUSIONTdmu:Source
  • Soluble Lectin-Like Oxidized LDL Receptor 1 as a Possible Mediator of Endothelial Dysfunction in Patients With Metabolic SyndromePubmed:24798146
  • Chemerin plays a protective role by regulating human umbilical vein endothelial cell-induced nitric oxide signaling in preeclampsiaPubmed:24840719
  • A Remarkable Age-Related Increase in SIRT1 Protein Expression against Oxidative Stress in Elderly:PubMed: 25785999
  • Comparative angioprotective effects of magnesium compoundsPubMed: 25127069
  • Peptides from hydrolysate of lantern fish (Benthosema pterotum) proved neuroprotective in vitro and in vivoscience:S1756464616300639
  • Роль индуцибельной NO-синтазы в механизмах регуляции функциональной активности кальций-активируемых калиевых каналов коронарных сосудов у крыс, адаптированных к стрессуarticle:n
  • Mutant DD genotype of NFKB1 gene is associated with the susceptibility and severity of coronary artery disease.pubmed:28088561
  • ThE REmOdELINg Of LEfT ATRIUm ANd vAsOdILATION fAcTORs chANgEs IN IschEmIc hEART fAILURE7038
  • Endothelial nitric oxide synthase deficiency influences normal cell cycle progression andapoptosis in trabecular meshwork cells.pubmed:27366677
  • SIRT1 inhibits releases of HMGB1 and HSP70 from human umbilical vein endothelial cells caused by IL-6 and the serum from a preeclampsia patient and protects the cells from death.pubmed:28122311
  • SIRT1 inhibits releases of HMGB1 and HSP70 from human umbilical vein endothelial cells caused by IL-6 and the serum from a preeclampsia patient and protects the cells from deathDOI:10.1016/j.biopha.2017.01.087
  • THE INFLUENCE OF THE ADDITIONAL USAGE OF L-ARGININE ON THE FACTORS OF VASODILATION AND THE CLINICAL DATA, COMPLICATED WITH CARDIAC DECOMPENSATION WITH REDUCED AND PRESERVED FRACTURE OF THE LEFT VENTRICLE THAT PATIENTS WITH STABLE CORONARY HEART DISEASE HAVEISSN: 0976-3376
  • Blunted Nitric Oxide Regulation in Tibetans under High Altitude Hypoxia10.1093:nsr
  • Molecular basis of vascular damage caused by cigarette smoke exposure and a new approach to the treatment: Alpha-linolenic acidPubmed:29579706
  • Ergothioneine prevents endothelial dysfunction induced by mercury chloridePubmed:29805489
  • Effect of statins on sirtuin 1 and endothelial nitric oxide synthase expression in young patients with a history of premature myocardial infarctionPubmed:29664427
  • Stężenia śródbłonkowej i indukowalnej formy syntazy tlenku azotu w surowicy krwi kobiet ciężarnych palących tytoń–badania wstępne
  • Tadalafil Preserves Penile Nitric Oxide Synthase from Detrimental Effect of Paroxetine in RatsDoi: 10.5152/eurasianjmed.2018.18160
  • Influence of Active Exposure to Tobacco Smoke on Nitric Oxide Status of Pregnant WomenPubmed: 30513890
  • Premature Myocardial Infarction: Genetic Variations in SIRT1 Affect Disease Susceptibility
  • Silica nanoparticles trigger the vascular endothelial dysfunction and prethrombotic state via miR-451 directly regulating the IL6R signaling pathwayPubmed: 30975181
  • ВЛИЯНИЕ ИНДУЦИБЕЛЬНОЙ NO-СИНТАЗЫ НА ТОНУС КОРОНАРНЫХ СОСУДОВ, СОКРАТИТЕЛЬНУЮ ФУНКЦИЮ МИОКАРДА И МЕХАНИЗМЫ …
  • Динамика показателей эндотелиальной дисфункции до и после операции у больных варикозной болезнью нижних конечностей
  • The placental growth factor attenuates intimal hyperplasia in vein grafts by improving endothelial dysfunctionPubmed: 31836533
  • Circadian fluctuations of endothelial nitric oxide synthase activity in females with rheumatoid arthritis: a pilot studyPubmed: 32025851
  • Circadian rhythms of endothelial nitric oxide synthase and toll-like receptors 2 production in females with rheumatoid arthritis depending on NOS3 gene polymorphism …Pubmed: 32297585
  • ASSOCIATIONS BETWEEN EFFICACY OF THE THERAPY AND CIRCADIAN FLUCTUATIONS OF ENDOTHELIAL NITRIC OXIDE SYNTHASE WITH TOLL-LIKE …Pubmed: 32242853
  • Dynamics of endothelial dysfunction indices before and after operation in patients with varicose disease of the lower extremities
  • Benzimidazole Derivative, 2-ETH Prevents Poststressor Disorders of Coronary Vascular Tone and Myocardial Contractility
  • Оксидативный стресс и эндотелиальная дисфункция в развитии экспериментального радиоиндуцированного повреждения легких и их коррекция …
  • Angiogenic factors measured in aspirated placental tissue between the 10+ 6 and 18+ 3 weeks of gestation34798517
  • Anti-atherosclerotic activity of aqueous extract of Ipomoea batatas (L.) leaves in high-fat diet-induced atherosclerosis model ratsPubmed:34986543
  • Serum inducible and endothelial nitric oxide synthase in coronary artery disease patients with Type 2 Diabetes mellitusPubmed:35647851
  • Mechanism of Fructus Mume Pills Underlying Their Protective Effects in Rats with Acetic Acid-Inducedulcerative Colitis via the Regulation of Inflammatory …Pubmed:35620404

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