High Sensitive ELISA Kit for Apolipoprotein B (APOB)

Apo-B; FLDB; Apo B-100; Apo B-48

Specificity

This assay has high sensitivity and excellent specificity for detection of High Sensitive Apolipoprotein B (APOB).
No significant cross-reactivity or interference between High Sensitive Apolipoprotein B (APOB) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 93-101 97
EDTA plasma(n=5) 79-101 83
heparin plasma(n=5) 81-90 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive Apolipoprotein B (APOB) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive Apolipoprotein B (APOB) 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 Apolipoprotein B (APOB) 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) 82-92% 80-96% 91-99% 83-91%
EDTA plasma(n=5) 86-93% 78-101% 82-102% 96-104%
heparin plasma(n=5) 89-96% 99-105% 80-104% 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 Lipid Research Group 1B phospholipase A2 deficiency protects against diet-induced hyperlipidemia in mice Jlr: M019463
J Clin Endocrinol Metab.? Elevated retinol binding protein 4 induces apolipoprotein B production and associates with hypertriglyceridemia Pubmed:25781360
Biochim Biophys Acta Culturing of HepG2 cells with human serum improve their functionality and suitability in studies of lipid metabolism PubMed: 26515253
Am J Cardiovasc Dis Hypoascorbemia induces atherosclerosis and vascular deposition of lipoprotein (a) in transgenic mice PubMed: 26064792
The Journal of Biological Chemistry Proprotein convertase subtilisin/Kexin type 9 (PCSK9) single domain antibodies are potent inhibitors of LDL receptor degradation Pubmed:27284008
United States Patent Application COMPOSITIONS AND METHODS FOR EFFICACIOUS AND SAFE DELIVERY OF SIRNA USING SPECIFIC CHITOSAN-BASED NANOCOMPLEXES y2016:0152988.html
alcohol and alcoholism Hops (Humulus lupulus) Content in Beer Modulates Effects of Beer on the Liver After Acute Ingestion in Female Mice. pubmed:27659607
Frontiers in Physiology Change in the Lipid Transport Capacity of the Liver and Blood during Reproduction in Rats pubmed:28798692
Food Research International Wild Lonicera caerulea berry polyphenol extract reduces cholesterol accumulation and enhances antioxidant capacity in vitro and in vivo Pubmed:29580541
Journal of The Science of Food and Agriculture Consumption of orange fermented beverage improves antioxidant status and reduces peroxidation lipid and inflammatory markers in healthy humans Pubmed:29124773
European Journal of Pharmacology The effect of exenatide (a GLP-1 analog) and sitagliptin (a DPP-4 inhibitor) on plasma platelet-activating factor acetylhydrolase (PAF-AH) activity and concentration in?… Pubmed: 30768981
Nature Communications Hepatic HuR modulates lipid homeostasis in response to high-fat diet Pubmed: 32546794
Study of in Vivo Serum Lipid Regulation with Ulmus macrocarpa Hance Extract in Rats
Nat Commun GP73 is a TBC-domain Rab GTPase-activating protein contributing to the pathogenesis of non-alcoholic fatty liver disease without obesity 34853313
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