Multiplex Assay Kit for Paraoxonase 1 (PON1) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Paraoxonase 1 (PON1) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Paraoxonase 1 (PON1) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-104 90
EDTA plasma(n=5) 79-105 88
heparin plasma(n=5) 98-105 102
sodium citrate plasma(n=5) 78-90 81

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Paraoxonase 1 (PON1) ,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 Paraoxonase 1 (PON1) ,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 Paraoxonase 1 (PON1) ,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) 89-98% 82-101% 93-101% 96-103%
EDTA plasma(n=5) 94-103% 84-96% 97-104% 83-94%
heparin plasma(n=5) 91-105% 83-91% 85-98% 96-105%
sodium citrate plasma(n=5) 98-105% 96-103% 96-105% 89-103%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:PON1) 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.

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Magazine Citations
Digestive Diseases and Sciences Fatty Liver Is Associated with Recurrent Bacterial Infections Independent of Metabolic Syndrome PubMed: 20205513
Clinica Chimica Acta Persistent elevation of paraoxonase-1 specific enzyme activity after weight reduction in obese non-diabetic men with metabolic syndrome PubMed: 21704607
Journal of Laparoendoscopic & Advanced Surgical Techniques Combined Serum Proteomic and Metabonomic Profiling After Laparoscopic Sleeve Gastrectomy in Children and Adolescents Wiley: source
PLoS One. High density lipoprotein structural changes and drug response in lipidomic profiles following the long-term fenofibrate therapy in the FIELD substudy PubMed: 21887280
Atherosclerosis Macrophage cholesterol efflux to plasma and HDL in subjects with low and high homocysteine levels: a FIELD substudy PubMed: 21696738
Lipids in Health and Disease Beneficial effects of omega-3 fatty acids in the proteome of high-density lipoprotein proteome PubMed: PMC3480889
British Journal of Nutrition Extra-virgin olive oil consumption reduces the age-related decrease in HDL and paraoxonase 1 anti-inflammatory activities Pubmed: 23510814
Diabetes & Metabolic Syndrome Paraoxonase 1: A better atherosclerotic risk predictor than HDL in type 2 diabetes mellitus Pubmed: 23680251
Redox Report Paraoxonase responses to exercise and niacin therapy in men with metabolic syndrome Pubmed:25180827
Toxicol Ind Health. Influence of α-tocopherol and α-lipoic acid on bisphenol-A-induced oxidative damage in liver and ovarian tissue of rats Pubmed:25548375
RETOS. Nuevas Tendencias en Educación Física, Deporte y Recreación A single session of aerobic exercise influences paraoxonase 1 activity and concentration Articulo: 345738764042
PLoS One Plasma Protein Biomarkers of Hepatocellular Carcinoma in HCV-Infected Alcoholic Patients with Cirrhosis PubMed: 25789864
Renal Failure Association of lipid profile with serum PON1 concentration in patients with chronic kidney disease Pubmed:26888625
Pharmacological reports The Role of PON-1, GR, IL-18, and oxLDL in Depression With and Without Posttraumatic Stress Disorder science:article
Clinical and Experimental Hypertension Association of paraoxonase 1 (PON1) gene polymorphisms and concentration with essential hypertension pubmed:27668323
Diabetes & Metabolic Syndrome: Clinical Research & Reviews Paraoxonase-1 is a better indicator than HDL of Atherosclerosis-A pilot Study in North Indian population pubmed:29254890
Diabetes/Metabolism Research and Reviews A Case-control Study: The Association of Serum Paraoxonase 1 Activity and Concentration with the Development of Type 2 Diabetes Mellitus pubmed:29156090
Journal of Chromatography A 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 stenosis S0021967317311858
Pharmacological Reports The role of PON-1, GR, IL-18, and OxLDL in depression with and without posttraumatic stress disorder. pubmed:28623707
American Journal of Food Science and Health Palm Wine and Orijin Bitters Severed Hyperlipidemia and Immunomodulatory Responses in Atherogenic-diet Fed Male Wistar Rats ISSN: 2381-7224
Metabolic brain disease Individuals with autism have higher 8-Iso-PGF2α levels than controls, but no correlation with quantitative assay of Paraoxonase 1 serum levels pubmed:28808839
Therapeutic Apheresis And Dialysis Serum Endocan Levels are Associated With Paraoxonase 1 Concentration in Patients With Chronic Kidney Disease Pubmed:29368426
Biological Trace Element Research Effect of Aqueous Extract of Glycyrrhiza glabra on the Biochemical Changes Induced by Cadmium Chloride in Rats Pubmed: 30251228
GLYCOCONJUGATE JOURNAL Identification a novel clinical biomarker in early diagnosis of human non-small cell lung cancer Pubmed: 30607521
Hematology. Differentially expressed plasma proteins of β-thalassemia/hemoglobin E patients in response to curcuminoids/vitamin E antioxidant cocktails Pubmed: 30661467
Proteomics Clinical Applications Comparative proteomics analysis of serum proteins in gestational diabetes during early and middle stages of pregnancy Pubmed: 31162828
Environment International Toxicity assessment due to prenatal and lactational exposure to lead, cadmium and mercury mixtures Pubmed: 31639605
Nutrition, Metabolism and Cardiovascular Diseases Phenotypes and concentration of PON1 in cardiovascular disease: the role of nutrient intake Pubmed: 31757567
Environment International Estimating renal and hepatic clearance rates of organophosphate esters in humans: Impacts of intrinsic metabolism and binding affinity with plasma proteins Pubmed: 31783242
RESPIRATORY RESEARCH The HDL from septic-ARDS patients with composition changes exacerbates pulmonary endothelial dysfunction and acute lung injury induced by cecal ligation … Pubmed: 33148285
CLINICAL RHEUMATOLOGY PON-1 haplotype (-108C> T, L55M, and Q192R) modulates the serum levels and activity PONase promoting an atherogenic lipid profile in rheumatoid arthritis patients Pubmed: 32556934
respiratory research The adverse alterations of HDL in quality promote septic ARDS via exacerbated pulmonary endothelial dysfunction
J Transl Med Multi-omic signatures of atherogenic dyslipidaemia: pre-clinical target identification and validation in humans 33407555
PLoS One Vitamin D status and systemic redox biomarkers in adults with obesity 30973911
Oxid Med Cell Longev Repetitions of Strenuous Exercise Consistently Increase Paraoxonase 1 Concentration and Activity in Plasma of Average-Trained Men 34917230
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