ELISA Kit for Paraoxonase 1 (PON1) Homo sapiens (Human) Sandwich ELISA

ESA; PON; Esterase A; Serum paraoxonase/arylesterase 1; Aromatic esterase 1; A-esterase 1; Serum aryldialkylphosphatase 1

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  • ELISA Kit for Paraoxonase 1 (PON1) Packages (Simulation)
  • ELISA Kit for Paraoxonase 1 (PON1) Packages (Simulation)
  • ELISA Kit for Paraoxonase 1 (PON1) Results demonstration
  • SEA243Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Paraoxonase 1 (PON1) and the recovery rates were calculated by comparing the measured value to the expected amount of Paraoxonase 1 (PON1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 89-97 93
EDTA plasma(n=5) 87-102 93
heparin plasma(n=5) 84-91 88

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Paraoxonase 1 (PON1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Paraoxonase 1 (PON1) 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 Paraoxonase 1 (PON1) 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) 89-98% 87-101% 84-98% 80-103%
EDTA plasma(n=5) 85-98% 78-93% 89-101% 97-105%
heparin plasma(n=5) 80-90% 80-91% 84-94% 88-101%

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 Paraoxonase 1 (PON1)

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 Paraoxonase 1 (PON1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Paraoxonase 1 (PON1). 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 Paraoxonase 1 (PON1), 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 Paraoxonase 1 (PON1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Fatty Liver Is Associated with Recurrent Bacterial Infections Independent of Metabolic SyndromePubMed: 20205513
  • Persistent elevation of paraoxonase-1 specific enzyme activity after weight reduction in obese non-diabetic men with metabolic syndromePubMed: 21704607
  • Combined Serum Proteomic and Metabonomic Profiling After Laparoscopic Sleeve Gastrectomy in Children and AdolescentsWiley: source
  • High density lipoprotein structural changes and drug response in lipidomic profiles following the long-term fenofibrate therapy in the FIELD substudyPubMed: 21887280
  • Macrophage cholesterol efflux to plasma and HDL in subjects with low and high homocysteine levels: a FIELD substudyPubMed: 21696738
  • Beneficial effects of omega-3 fatty acids in the proteome of high-density lipoprotein proteomePubMed: PMC3480889
  • Extra-virgin olive oil consumption reduces the age-related decrease in HDL and paraoxonase 1 anti-inflammatory activitiesPubmed: 23510814
  • Paraoxonase 1: A better atherosclerotic risk predictor than HDL in type 2 diabetes mellitusPubmed: 23680251
  • Paraoxonase responses to exercise and niacin therapy in men with metabolic syndromePubmed:25180827
  • Influence of α-tocopherol and α-lipoic acid on bisphenol-A-induced oxidative damage in liver and ovarian tissue of ratsPubmed:25548375
  • A single session of aerobic exercise influences paraoxonase 1 activity and concentrationArticulo: 345738764042
  • Plasma Protein Biomarkers of Hepatocellular Carcinoma in HCV-Infected Alcoholic Patients with CirrhosisPubMed: 25789864
  • Association of lipid profile with serum PON1 concentration in patients with chronic kidney diseasePubmed:26888625
  • The Role of PON-1, GR, IL-18, and oxLDL in Depression With and Without Posttraumatic Stress Disorderscience:article
  • Association of paraoxonase 1 (PON1) gene polymorphisms and concentration with essential hypertensionpubmed:27668323
  • Paraoxonase-1 is a better indicator than HDL of Atherosclerosis-A pilot Study in North Indian populationpubmed:29254890
  • A Case-control Study: The Association of Serum Paraoxonase 1 Activity and Concentration with the Development of Type 2 Diabetes Mellituspubmed:29156090
  • Application of a new procedure for liquid chromatography/mass spectrometry profiling of plasma amino acid-related metabolites and untargeted shotgun proteomics to identify mechanisms and biomarkers of calcific aortic stenosisS0021967317311858
  • The role of PON-1, GR, IL-18, and OxLDL in depression with and without posttraumatic stress disorder.pubmed:28623707
  • Palm Wine and Orijin Bitters Severed Hyperlipidemia and Immunomodulatory Responses in Atherogenic-diet Fed Male Wistar RatsISSN: 2381-7224
  • Individuals with autism have higher 8-Iso-PGF2α levels than controls, but no correlation with quantitative assay of Paraoxonase 1 serum levelspubmed:28808839
  • Serum Endocan Levels are Associated With Paraoxonase 1 Concentration in Patients With Chronic Kidney DiseasePubmed:29368426
  • Effect of Aqueous Extract of Glycyrrhiza glabra on the Biochemical Changes Induced by Cadmium Chloride in RatsPubmed: 30251228
  • Identification a novel clinical biomarker in early diagnosis of human non-small cell lung cancerPubmed: 30607521
  • Differentially expressed plasma proteins of β-thalassemia/hemoglobin E patients in response to curcuminoids/vitamin E antioxidant cocktailsPubmed: 30661467
  • Comparative proteomics analysis of serum proteins in gestational diabetes during early and middle stages of pregnancyPubmed: 31162828
  • Toxicity assessment due to prenatal and lactational exposure to lead, cadmium and mercury mixturesPubmed: 31639605
  • Phenotypes and concentration of PON1 in cardiovascular disease: the role of nutrient intakePubmed: 31757567
  • Estimating renal and hepatic clearance rates of organophosphate esters in humans: Impacts of intrinsic metabolism and binding affinity with plasma proteinsPubmed: 31783242
  • The HDL from septic-ARDS patients with composition changes exacerbates pulmonary endothelial dysfunction and acute lung injury induced by cecal ligation …Pubmed: 33148285
  • PON-1 haplotype (-108C> T, L55M, and Q192R) modulates the serum levels and activity PONase promoting an atherogenic lipid profile in rheumatoid arthritis patientsPubmed: 32556934
  • The adverse alterations of HDL in quality promote septic ARDS via exacerbated pulmonary endothelial dysfunction
  • Multi-omic signatures of atherogenic dyslipidaemia: pre-clinical target identification and validation in humans33407555
  • Vitamin D status and systemic redox biomarkers in adults with obesity30973911
  • Repetitions of Strenuous Exercise Consistently Increase Paraoxonase 1 Concentration and Activity in Plasma of Average-Trained Men34917230

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