Multiplex Assay Kit for Apolipoprotein A1 (APOA1) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

(Note: Up to 8-plex in one testing reaction)

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 98-105 101
EDTA plasma(n=5) 80-95 91
heparin plasma(n=5) 96-104 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Apolipoprotein A1 (APOA1) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 98-105% 86-93% 81-97% 93-104%
EDTA plasma(n=5) 80-101% 96-104% 81-99% 86-103%
heparin plasma(n=5) 79-103% 94-102% 88-96% 89-97%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:APOA1) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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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
ACS Biomaterials Science & Engineering Enhanced Antiatherosclerotic Efficacy of Statin-Loaded Reconstituted High-Density Lipoprotein via Ganglioside GM1 Modification abs:10.1021
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
Toxicon A membrane disrupting toxin from wasp venom underlies the molecular mechanism of tissue damage Pubmed:29654869
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
European Journal of Preventive Cardiology High-intensity interval training has a greater effect on reverse cholesterol transport elements compared with moderate-intensity continuous training in obese male rats Pubmed: 31718266
Diagnostics Systemic Alterations of Immune Response-Related Proteins during Glaucoma Development in the Murine Model DBA/2J Pubmed: 32585848
VETERINARY PATHOLOGY Pathology and Proteomics-Based Diagnosis of Localized Light-Chain Amyloidosis in Dogs and Cats Pubmed: 32880234
Nat Commun GP73 is a TBC-domain Rab GTPase-activating protein contributing to the pathogenesis of non-alcoholic fatty liver disease without obesity 34853313
Cardiovasc Toxicol Interplay of Obesity, Ethanol, and Contaminant Mixture on Clinical Profiles of Cardiovascular and Metabolic Diseases: Evidence from an Animal Study Pubmed:35429258
Peptides Neuropeptide Y promotes hepatic apolipoprotein A1 synthesis and secretion through neuropeptide Y Y5 receptor Pubmed:35660638
Journal of Functional Foods Castanea mollissima shell polyphenols regulate JAK2 and PPARγ expression to suppress inflammation and lipid accumulation by inhibiting M1 macrophages …
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