ELISA Kit for Apolipoprotein A1 (APOA1)

Apo-A1; ApoA-1 Milano; ProapoA-I; Proapolipoprotein A-I; Truncated apolipoprotein A-I

Specificity

This assay has high sensitivity and excellent specificity for detection of Apolipoprotein A1 (APOA1).
No significant cross-reactivity or interference between Apolipoprotein A1 (APOA1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Apolipoprotein A1 (APOA1) and the recovery rates were calculated by comparing the measured value to the expected amount of Apolipoprotein A1 (APOA1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 93-101 96
EDTA plasma(n=5) 82-89 85
heparin plasma(n=5) 91-105 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Apolipoprotein A1 (APOA1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Apolipoprotein A1 (APOA1) 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 Apolipoprotein A1 (APOA1) 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) 78-96% 86-101% 91-99% 95-104%
EDTA plasma(n=5) 97-104% 97-105% 87-103% 88-95%
heparin plasma(n=5) 80-99% 96-105% 94-102% 93-105%

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
European Journal of Applied Physiology Endurance training enhances ABCA1 expression in rat small intestine PubMed: 19629515
Official Journal of the INWR Effect of eight weeks of wrestling and circuit fitness training on Apo lipoportein A-I and lymphocyte ABCA1 gene expression in well-trained wrestler. Wrestling: source
Biochemical and Biophysical Research Communications Treadmill exercise enhances ABCA1 expression in rat liver ScienceDirect: S0006291X0701488X
World Journal of Sport Sciences The Effect of Eight Weeks of Wrestling and Wrestling Technique Based Circuit Training on Lymphocyte ABCA1 Gene Expression and Plasma Apolipoprotein A-1 Idosi: source
Biochimica et Biophysica Acta Hepatic lipase- and endothelial lipase-deficiency in mice promotes macrophage-to-feces RCT and HDL antioxidant properties PubMed: 23328279
Biomarkers The association of a distinct plasma proteomic profile with the cervical high-grade squamous intraepithelial lesion of Uyghur women: a 2D liquid-phase chromatography/mass spectrometry study PubMed: 22458349
Journal of the American Heart Association MicroRNA-33 Deficiency Reduces the Progression of Atherosclerotic Plaque in ApoE?/? Mice PubMed: PMC3540673
PLoS ONE Apolipoprotein AI Is a Potential Mediator of Remote Ischemic Preconditioning Plosone: Source
Molecular Genetics and Metabolism DYRK1A overexpression decreases plasma lecithin: cholesterol acyltransferase activity and apolipoprotein AI levels Pubmed: 23920041
Endocrinology Deficiency of Clusterin Exacerbates High-Fat Diet-Induced Insulin Resistance in Male Mice Pubmed: 24684302
Molecular Genetics and Metabolism Reports Corrective effects of hepatotoxicity by hepatic Dyrk1a gene delivery in mice with intermediate hyperhomocysteinemia Sciencedirect:S2214426915000026
J Psychopharmacol Anti-atherogenic properties of high-density lipoproteins in psychiatric patients before and after two months of atypical anti-psychotic therapy PubMed: 26253619
Biochim Biophys Acta Culturing of HepG2 cells with human serum improve their functionality and suitability in studies of lipid metabolism PubMed: 26515253
Pharmacol Res Perspect Efficacy of tomato concentrates in mouse models of dyslipidemia and cancer PubMed: 26171234
Clin Exp Nephrol Differentially expressed urinary biomarkers in children with idiopathic nephrotic syndrome PubMed: 26351173
Oncogene Apolipoprotein A-I inhibits experimental colitis and colitis-propelled carcinogenesis PubMed: 26279300
Oncogene Apolipoprotein AI inhibits experimental colitis and colitis-propelled carcinogenesis Pubmed:26279300
Allergy Serum levels of 9α,11β-PGF2 and apolipoprotein A1 achieve high predictive power as biomarkers of anaphylaxis. pubmed:28378321
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
11 High-Density Lipoproteins Exert Pro-inflammatory Effects on Macrophages via Passive Cholesterol Depletion and PKC-NF-kB/STAT1-IRF1 Signaling pubmed:27866837
Biomedical Science Letters Effect of Ginkgo biloba Extract (EGb 761) on Serum Cholesterol Levels in Wild-type C57Bl/6 Mice 10.15616/BSL.2017.23.2.80
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