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) 87-96 92
EDTA plasma(n=5) 99-105 102
heparin plasma(n=5) 79-103 101

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) 78-102% 88-98% 83-93% 90-97%
EDTA plasma(n=5) 92-102% 90-104% 90-99% 91-99%
heparin plasma(n=5) 85-103% 92-105% 88-95% 90-97%

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.

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Magazine Citations
Journal of Ethnopharmacology Total panax notoginsenosides prevent atherosclerosis in apolipoprotein E-knockout mice: Role of downregulation of CD40 and MMP-9 expression ScienceDirect: S0378874109005091
Autonomic Neuroscience Chemical sympathectomy induces arterial accumulation of native and oxidized LDL in hypercholesterolemic rats ScienceDirect: S1566070211004152
Cell Biochemistry and Biophysics Plasma Paraoxonase-1, Oxidized Low-Density Lipoprotein and Lipid Peroxidation Levels in Gout Patients SpringerLink: u466114p03434m6t
Cardiovascular Research Endothelial-specific Nox2 overexpression increases vascular superoxide and macrophage recruitment in ApoE-/- mice PubMed: PMC3307381
Atherosclerosis 10-Dehydrogingerdione raises HDL-cholesterol through a CETP inhibition and wards off oxidation and inflammation in dyslipidemic rabbits. Pubmed: 24267247
PloS one Atheroprotective effect of oleoylethanolamide (OEA) targeting oxidized LDL. NCBI: PMC3896367
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J Surg Arts THE FUNCTION OF SIALIC ACID AS A RADICAL SCAVENGER IN EXPERIMENTAL HYPOTHYROIDISM WITH AND WITHOUT HYPERLIPIDEMIA. Cerrahisanatlardergisi:Source
PLoS One. Atheroprotective effect of oleoylethanolamide (OEA) targeting oxidized LDL Pubmed:24465540
Saudi Med J Increased systemic low-grade inflammation in high altitude native rats mediated by adrenergic receptors Pubmed:24888651
Vascular Medicine Anti‐Inflammatory Immune Skewing Is Atheroprotective: Apoe−/−FcγRIIb−/− Mice Develop Fibrous Carotid Plaques Pubmed:25516435
Journal of Functional Foods Oral administration of the β-glucan produced by Aureobasidium pullulans ameliorates development of atherosclerosis in apolipoprotein E deficient mice Science: Article
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
BMJ Open Sport Exerc Med Athletes with higher VO2max present reduced oxLDL after a marathon race Content: 1
Biochim Biophys Acta PPARβ/δ ameliorates fructose-induced insulin resistance in adipocytes by preventing Nrf2 activation PubMed: 25728706
PLoS One Ezetimibe Attenuates Atherosclerosis Associated with Lipid Reduction and Inflammation Inhibition PubMed: 26555472
PLoS One Pulsatile Hyperglycaemia Induces Vascular Oxidative Stress and GLUT 1 Expression More Potently than Sustained Hyperglycaemia in Rats on High Fat Diet Pubmed:26790104
Diabetology & Metabolic Syndrome Oxidative DNA damage and oxidized low density lipoprotein in Type II diabetes mellitus among patients with Helicobacter pylori infection pmc:PMC4855822
J Am Heart Assoc. Transcriptional Profiling of Foam Cells Reveals Induction of Cavia (Guinea pig )anylate‐Binding Proteins Following Western Diet Acceleration of Atherosclerosis in the Absence of Global Changes in Inflammation pmc:PMC4859273
Journal of Applied Pharmaceutical Science Organic Arsenical Exposure Stimulates Atherosclerosis through Oxidative Stress Increase and Adhesion Molecule Expression admin:php
Journal of Nutritional Biochemistry Dietary rose hip exerts anti-atherosclerotic effects and increases nitric oxide-mediated dilation in ApoE-null mice S0955286316301887
Journal of Basic and Clinical Physiology and Pharmacology Effects of altered thyroid states on oxidative stress parameters in rats pubmed:27824612
Journal of Functional Foods Atherosclerosis-related inflammation and oxidative stress are improved by rice bran enzymatic extract S1756464616302511
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