Multiplex Assay Kit for Endothelial NOS (eNOS) ,etc. by FLIA (Flow Luminescence Immunoassay) Rattus norvegicus (Rat) Multiplex ELISA

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

(Note: Up to 8-plex in one testing reaction)

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  • Multiplex Assay Kit for Endothelial NOS (eNOS) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Endothelial NOS (eNOS) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Endothelial NOS (eNOS) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA868Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 81-97 86
EDTA plasma(n=5) 86-101 94
heparin plasma(n=5) 91-99 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Endothelial NOS (eNOS) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 78-97% 96-104% 82-101% 96-105%
EDTA plasma(n=5) 83-102% 93-101% 96-103% 85-93%
heparin plasma(n=5) 82-93% 78-97% 78-102% 78-104%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:eNOS) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

Multiplex Assay Kit for Endothelial NOS (eNOS) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer.The MFI developed is proportional to the concentration of analytes of interest in the sample.

Giveaways

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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