Multiplex Assay Kit for Angiopoietin Like Protein 2 (ANGPTL2) ,etc. by CBA (Cytometric Bead Array)

HARP; ARP2; Angiopoietin-Related Protein 2

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 90-104 97
EDTA plasma(n=5) 78-105 99
heparin plasma(n=5) 82-98 87

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Angiopoietin Like Protein 2 (ANGPTL2) ,etc. by CBA (Cytometric Bead Array) 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) ,etc. by CBA (Cytometric Bead Array) 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) ,etc. by CBA (Cytometric Bead Array) 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) 91-98% 80-104% 80-99% 87-96%
EDTA plasma(n=5) 89-97% 84-97% 80-105% 92-105%
heparin plasma(n=5) 95-104% 92-105% 80-90% 90-103%

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

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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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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