ELISA Kit for Angiopoietin Like Protein 4 (ANGPTL4)

ARP4; FIAF; HFARP; NL2; PGAR; Pp1158; Fasting-Induced Adipose Factor; Hepatic Angiopoietin-Related Protein; Hepatic Fibrinogen/Angiopoietin-Related Protein

Specificity

This assay has high sensitivity and excellent specificity for detection of Angiopoietin Like Protein 4 (ANGPTL4).
No significant cross-reactivity or interference between Angiopoietin Like Protein 4 (ANGPTL4) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Angiopoietin Like Protein 4 (ANGPTL4) and the recovery rates were calculated by comparing the measured value to the expected amount of Angiopoietin Like Protein 4 (ANGPTL4) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-98 87
EDTA plasma(n=5) 96-103 99
heparin plasma(n=5) 81-95 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Angiopoietin Like Protein 4 (ANGPTL4) 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 4 (ANGPTL4) 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 4 (ANGPTL4) 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) 85-96% 79-92% 78-104% 91-99%
EDTA plasma(n=5) 91-98% 95-104% 94-104% 85-102%
heparin plasma(n=5) 94-101% 96-104% 83-93% 79-89%

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
The Journal of Biological Chemistry Angiopoietin-like 4 (Angptl4) Protein Is a Physiological Mediator of Intracellular Lipolysis in Murine Adipocytes PubMed: PMC3318686
J Clin Endocrinol Metab. Decreased Concentrations of the Lipoprotein Lipase Inhibitor Angiopoietin-Like Protein 4 and Increased Serum Triacylglycerol Are Associated With Increased Neonatal Fat Mass in Pregnant Women With Gestational Diabetes Mellitus Pubmed: 23744407
PloS one Lack of Relationship between Cord Serum Angiopoietin-Like Protein 4 (ANGPTL4) and Lipolytic Activity in Human Neonates Born by Spontaneous Delivery Pubmed: 24324678
Journal of Hematology & Oncology Angptl4 is upregulated under inflammatory conditions in the bone marrow of mice, expands myeloid progenitors, and accelerates reconstitution of platelets after myelosuppressive therapy PubMed: 26054961
Gynecol Endocrinol Examination of angiopoietin-like protein 4, neuropeptide Y, omentin-1 levels of obese and non-obese patients with polycystic ovary syndrome PubMed: 26291814
Molecular Nutrition & Food research Interaction of Gut Microbiota and High‐sodium, Low‐potassium Diet in Altering Plasma Triglyceride Profiles Revealed by Lipidomics Analysis Pubmed: 31675161
ANNALS OF AGRICULTURAL AND ENVIRONMENTAL MEDICINE Analysis of novel serum markers of fibrosis and angiogenesis in patients with alcoholic liver cirrhosis
Food Chem Toxicol Di (2-ethylhexyl) phthalate exposure exacerbates metabolic disorders in diet-induced obese mice 34303773
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