ELISA Kit for Angiopoietin Like Protein 2 (ANGPTL2)

HARP; ARP2; Angiopoietin-Related Protein 2

Specificity

This assay has high sensitivity and excellent specificity for detection of Angiopoietin Like Protein 2 (ANGPTL2).
No significant cross-reactivity or interference between Angiopoietin Like Protein 2 (ANGPTL2) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Angiopoietin Like Protein 2 (ANGPTL2) and the recovery rates were calculated by comparing the measured value to the expected amount of Angiopoietin Like Protein 2 (ANGPTL2) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-90 87
EDTA plasma(n=5) 83-101 91
heparin plasma(n=5) 89-97 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Angiopoietin Like Protein 2 (ANGPTL2) 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 2 (ANGPTL2) 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 2 (ANGPTL2) 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) 85-105% 93-105% 84-102% 87-104%
EDTA plasma(n=5) 94-103% 90-98% 83-93% 90-105%
heparin plasma(n=5) 87-103% 99-105% 80-103% 80-97%

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
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
Molecular and Cellular Endocrinology Tumor necrosis factor-α increases angiopoietin-like protein 2 gene expression by activating Foxo1 in 3T3-L1 adipocytes PubMed: 21501655
Journal of Leukocyte Biology Prolyl-4-hydroxylase domain 3 (PHD3) is a critical terminator for cell survival of macrophages under stress conditions Pubmed:24626957
J Am Heart Assoc. Lack of Angiopoietin‐Like‐2 Expression Limits the Metabolic Stress Induced by a High‐Fat Diet and Maintains Endothelial Function in Mice Pubmed:25128474
Clinical Cancer Research Elevated Serum Angiopoietin-like Protein 2 Correlates with the Metastatic Properties of Colorectal Cancer: A Serum Biomarker for Early Diagnosis and Recurrence Pubmed:25294915
Carcinogenesis Serum angiopoietin-like protein 2 as a potential biomarker for diagnosis, early recurrence and prognosis in gastric cancer patients PubMed: 26420253
Diabetologia ANGPTL2 is associated with an increased risk of cardiovascular events and death in diabetic patients. pubmed:27491833
Stem Cell Research and Therapy c-kit Positive Cardiac Outgrowth Cells Demonstrate Better Ability for Cardiac Recovery Against Ischemic Myopathy. pubmed:29238626
Cardiovasc Diabetol Influence of micro-and macro-vascular disease and Tumor Necrosis Factor Receptor 1 on the level of lower-extremity amputation in patients with type 2 … Pubmed:29879997
Biomedical Reports Circulating angiopoietin‑like protein 2 levels are associated with decreased renal function in HIV+ subjects on cART: A potential marker of kidney disease
Diabetologia Serum tenascin-C is independently associated with increased major adverse cardiovascular events and death in individuals with type 2 diabetes: a French … Pubmed: 32040670
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