Multiplex Assay Kit for Angiopoietin Like Protein 8 (ANGPTL8) ,etc. by FLIA (Flow Luminescence Immunoassay)

RIFL; TD26; C19orf80; Lipasin; Betatrophin; Hepatocellular Carcinoma-Associated Protein TD26

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Angiopoietin Like Protein 8 (ANGPTL8) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Angiopoietin Like Protein 8 (ANGPTL8) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Angiopoietin Like Protein 8 (ANGPTL8) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Angiopoietin Like Protein 8 (ANGPTL8) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 85-103 99
EDTA plasma(n=5) 91-99 95
heparin plasma(n=5) 84-104 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Angiopoietin Like Protein 8 (ANGPTL8) ,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 Angiopoietin Like Protein 8 (ANGPTL8) ,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 Angiopoietin Like Protein 8 (ANGPTL8) ,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) 87-98% 85-98% 81-98% 95-103%
EDTA plasma(n=5) 84-104% 90-101% 80-96% 98-105%
heparin plasma(n=5) 84-102% 93-105% 78-97% 89-101%

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#:ANGPTL8) 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
PLoS One Increased Maternal and Cord Blood Betatrophin in Gestational Diabetes PubMed: 26115519
Nutrición Hospitalaria Serum betatrophin level increased in subjects with metabolic syndrome: a case-control study wp-content:10563
Mediators of Inflammation Serum betatrophin level increased in subjects with nonalcoholic fatty liver disease files:ijcem0020951.pdf
PLoS One Resolving Discrepant Findings on ANGPTL8 in β-Cell Proliferation: A Collaborative Approach to Resolving the Betatrophin Controversy pubmed:27410263
Obesity Research and Clinical Practice Relationship between serum betatrophin levels and the first-phase of glucose-stimulated insulin secretion pubmed:27554132
打不开网页 Aumento del nivel de betatrofina en suero en sujetos con síndrome metabólico: estudio de casos y controles :
DIABETES & METABOLISM Levels of betatrophin decrease during pregnancy despite increased insulin resistance, beta-cellfunction and triglyceride levels. pubmed:27555469
Decreased circulating levels of betatrophin in Chinese women with polycystic ovary syndrome :
International Journal of Endocrinology Serum concentrations of betatrophin and its association with indirect indices of insulin resistance and beta cell function in women with polycystic ovary … pubmed:28702052
Diabetes Research and Clinical Practice Angiopoietin-like protein 8: an attractive biomarker for the evaluation of subjects with insulin resistance and related disorders pubmed:28965028
Diabetes/Metabolism Research and Reviews ANGPTL8 (Betatrophin) Role in Diabetes and Metabolic Diseases pubmed:28722798
Bibliography The Correlation Between Circulating Betatrophin and Insulin Resistance in General Population: A Meta-Analysis pubmed:28931172
Frontiers in Endocrinology ANGPTL8: an important regulator in metabolic disorders Pubmed:29719529
Nutrients ChREBP Reciprocally Regulates Liver and Plasma Triacylglycerol Levels in Different Manners Pubmed: 30405056
Nature Communications Angptl8 mediates food-driven resetting of hepatic circadian clock in mice Pubmed: 31388006
Journal of Immunology Research The Impact of Psoriasis and Metabolic Syndrome on the Systemic Inflammation and Oxidative Damage to Nucleic Acids Pubmed: 32537470
JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM Genetic and Metabolic Determinants of Plasma Levels of ANGPTL8 33619548
Medicine Association of betatrophin amounts with 25-(OH) D levels in patients with gestational diabetes mellitus 33879746
International Journal of Endocrinology Effect of Ileal Transposition (IT) on Angiopoietin-Like Protein-8 (ANGPTL8) and Pentraxin (PTX3) Plasma Level in Sprague-Dawley Rats Fed High-Fat Diet?¡­ 34035808
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