ELISA Kit for Oxidized Low Density Lipoprotein (OxLDL)

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Specificity

This assay has high sensitivity and excellent specificity for detection of Oxidized Low Density Lipoprotein (OxLDL).
No significant cross-reactivity or interference between Oxidized Low Density Lipoprotein (OxLDL) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Oxidized Low Density Lipoprotein (OxLDL) and the recovery rates were calculated by comparing the measured value to the expected amount of Oxidized Low Density Lipoprotein (OxLDL) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 87-97 92
EDTA plasma(n=5) 99-105 102
heparin plasma(n=5) 78-89 85

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Oxidized Low Density Lipoprotein (OxLDL) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Oxidized Low Density Lipoprotein (OxLDL) 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 Oxidized Low Density Lipoprotein (OxLDL) 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) 87-94% 85-98% 85-95% 92-105%
EDTA plasma(n=5) 95-102% 98-105% 95-105% 79-102%
heparin plasma(n=5) 91-105% 93-101% 91-104% 90-104%

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
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 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
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Autonomic Neuroscience Chemical sympathectomy induces arterial accumulation of native and oxidized LDL in hypercholesterolemic rats ScienceDirect: S1566070211004152
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Acta Pharmacologica Sinica Sirt1 is involved in decreased bone formation in aged apolipoprotein E-deficient mice PubMed: 26592520
Cancer Science Suppressive effects of the NADPH oxidase inhibitor apocynin on intestinal tumorigenesis in obese KK‐Ay and Apc mutant Min mice PubMed: 26310859
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PLoS One Ezetimibe Attenuates Atherosclerosis Associated with Lipid Reduction and Inflammation Inhibition PubMed: 26555472
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Journal of Nutritional Biochemistry Dietary rose hip exerts anti-atherosclerotic effects and increases nitric oxide-mediated dilation in ApoE-null mice S0955286316301887
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Nutrients Yirui Capsules Alleviate Atherosclerosis by Improving the Lipid Profile and Reducing Inflammation in Apolipoprotein E-Deficient Mice Pubmed:29382111
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Journal of Functional Foods Saponins of sea cucumber attenuate atherosclerosis in ApoE−/− mice via lipid-lowering and anti-inflammatory properties 10.1016:j.jff.2018.07.046
Journal of Agricultural and Food Chemistry Neuroprotective effect of hydroxytyrosol in experimental diabetic retinopathy: relationship with cardiovascular biomarkers Pubmed:29278909
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Lipids in Health and Disease Lipingshu capsule improves atherosclerosis associated with lipid regulation and inflammation inhibition in apolipoprotein E–deficient mice Pubmed:30064511
Diabetes Metabolism Research and Reviews  Diabetic dyslipidaemia is associated with alterations in eNOS, caveolin‐1, and endothelial dysfunction in streptozotocin treated rats Pubmed:29471582
Diabetes & Metabolic Syndrome: Clinical Research & Reviews Beyond LDL-c: The importance of serum oxidized LDL in predicting risk for type 2 diabetes in the middle-aged Asian Indians Doi: 10.1016/j.dsx.2018.08.036
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Document Version
MEDICINE Screening of schizophrenia associated miRNAs and the regulation of miR-320a-3p on integrin β1 Pubmed: 30813134
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Nature Communications ER-residential Nogo-B accelerates NAFLD-associated HCC mediated by metabolic reprogramming of oxLDL lipophagy Pubmed: 31358770
Clinical and Experimental Medicine Senescence-associated miR-34a and miR-126 in middle-aged Indians with type 2 diabetes Pubmed: 31732824
Steroids Serum 25 (OH) D concentration and its association with inflammation and oxidative stress in the middle-aged Indian healthy and diabetic subjects Pubmed: 31672627
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY Zygophyllum album saponins prevent atherogenic effect induced by deltamethrin via attenuating arterial accumulation of native and oxidized LDL in rats Pubmed: 32105945
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Journal of Diabetes Research Middle-Aged Indians with Type 2 Diabetes Are at Higher Risk of Biological Ageing with Special Reference to Serum CDKN2A Pubmed: 32280716
Acta Biochim Pol Dietary Lactobacillus plantarum LS/07 and inulin in the management of chronic disease risk factors Pubmed: 33108141
Ecotoxicol Environ Saf Permethrin induced arterial retention of native and oxidized LDL in rats by promoting inflammation, oxidative stress and affecting LDL receptors, and collagen … Pubmed: 32911180
Cell Death & Disease Oxidized-LDL inhibits testosterone biosynthesis by affecting mitochondrial function and the p38 MAPK/COX-2 signaling pathway in Leydig cells Pubmed: 32796811
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Eur J Nutr The effects of long-term moderate exercise and Western-type diet on oxidative/nitrosative stress, serum lipids and cytokines in female Sprague Dawley rats 34319428
Pediatr Gastroenterol Hepatol Nutr The Relationship between Daily Fructose Consumption and Oxidized Low-Density Lipoprotein and Low-Density Lipoprotein Particle Size in Children with … 34557400
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