Mini Samples ELISA Kit for Angiopoietin Like Protein 8 (ANGPTL8)

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Mini Samples Angiopoietin Like Protein 8 (ANGPTL8).
No significant cross-reactivity or interference between Mini Samples Angiopoietin Like Protein 8 (ANGPTL8) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 88-101 97
EDTA plasma(n=5) 90-98 93
heparin plasma(n=5) 90-103 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mini Samples Angiopoietin Like Protein 8 (ANGPTL8) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mini Samples Angiopoietin Like Protein 8 (ANGPTL8) 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 Mini Samples Angiopoietin Like Protein 8 (ANGPTL8) 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) 78-97% 98-105% 85-105% 83-90%
EDTA plasma(n=5) 79-88% 78-90% 92-104% 98-105%
heparin plasma(n=5) 98-105% 85-97% 95-104% 93-104%

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 1×60µL Assay Diluent A 1×6mL
Detection Reagent B 1×60µL Assay Diluent B 1×6mL
TMB Substrate 1×4.5mL Stop Solution 1×3mL
Wash Buffer (30 × concentrate) 1×10mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 25µL standard or sample to each well. Incubate 1 hour at 37°C;
3. Aspirate and add 25µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 25µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 25µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 20µL Stop Solution. Read at 450nm immediately.

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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
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