High Sensitive ELISA Kit for Low Density Lipoprotein (LDL)

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 92-101 96
EDTA plasma(n=5) 80-93 88
heparin plasma(n=5) 82-104 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive Low Density Lipoprotein (LDL) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive Low Density Lipoprotein (LDL) 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 High Sensitive Low Density Lipoprotein (LDL) 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) 79-94% 85-105% 94-103% 91-99%
EDTA plasma(n=5) 89-99% 92-105% 82-98% 80-89%
heparin plasma(n=5) 90-104% 82-101% 95-105% 82-94%

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
PloS one Atheroprotective effect of oleoylethanolamide (OEA) targeting oxidized LDL. NCBI: PMC3896367
PLoS One. Atheroprotective effect of oleoylethanolamide (OEA) targeting oxidized LDL Pubmed:24465540
J Lipid Res PCSK9 deficiency unmasks a sex-and tissue-specific subcellular distribution of the LDL and VLDL receptors in mice PubMed: 26323289
Neuroscience 27-Hydroxycholesterol contributes to disruptive effects on learning and memory by modulating cholesterol metabolism in the rat brain PubMed: 25987203
The Journal of Nutritional Biochemistry Protective role of n6/n3 PUFA supplementation with varying DHA/EPA ratios against atherosclerosis in mice Pubmed:27142749
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Int J Mol Sci. Combined Effects of Curcumin and Lycopene or Bixin in Yoghurt on Inhibition of LDL Oxidation and Increases in HDL and Paraoxonase Levels in Streptozotocin-Diabetic Rats. pubmed:28333071
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Journal of Functional Foods Red paprika (Capsicum annuum L.) and its main carotenoid capsanthin ameliorate impaired lipid metabolism in the liver and adipose tissue of high-fat diet-induced obese mice 10.1016:j.jff.2017.01.044
Microvascular Research Effects of insulin analogs as an add-on to metformin on cutaneous microcirculation in type 2 diabetic patients pubmed:28954218
Biochemical and Biophysical Research Communications Oxidized low-density lipoprotein promotes osteoclast differentiation from CD68 positive mononuclear cells by regulating HMGB1 release pubmed:29146189
Journal of Nutritional Science and Vitaminology (Tokyo) Anthocyanin Cyanidin-3-Glucoside Attenuates Platelet Granule Release in Mice Fed High-Fat Diets. pubmed:28978870
Environmental Science and Pollution Research Bifenthrin exerts proatherogenic effects via arterial accumulation of native and oxidized LDL in rats: the beneficial role of vitamin E and selenium Pubmed: 30465240
American Journal of Translational Research Alteration of circulating innate lymphoid cells in patients with atherosclerotic cerebral infarction Pubmed: 30662674
Nature Communications ER-residential Nogo-B accelerates NAFLD-associated HCC mediated by metabolic reprogramming of oxLDL lipophagy Pubmed: 31358770
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Evid Based Complement Alternat Med Effect of Acupoint Embedding on Serum Leptin and Hypothalamus Leptin Receptor Expression in Rats with Simple Obesity 34659432
Ecotoxicol Environ Saf High-fat diet-induced aggravation of cardiovascular impairment in permethrin-treated Wistar rats 34224971
Int J Food Sci Nutr Brown and golden flaxseed reduce intestinal permeability and endotoxemia, and improve the lipid profile in perimenopausal overweight women Pubmed:35311432
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