CLIA Kit for Angiopoietin 2 (ANGPT2)

ANG2; AGPT2

Specificity

This assay has high sensitivity and excellent specificity for detection of Angiopoietin 2 (ANGPT2).
No significant cross-reactivity or interference between Angiopoietin 2 (ANGPT2) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Angiopoietin 2 (ANGPT2) and the recovery rates were calculated by comparing the measured value to the expected amount of Angiopoietin 2 (ANGPT2) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 91-99 96
EDTA plasma(n=5) 82-90 85
heparin plasma(n=5) 98-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Angiopoietin 2 (ANGPT2) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Angiopoietin 2 (ANGPT2) 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 2 (ANGPT2) 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) 87-104% 80-92% 80-94% 97-105%
EDTA plasma(n=5) 86-101% 80-104% 95-103% 93-101%
heparin plasma(n=5) 86-97% 88-98% 83-105% 98-105%

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
Substrate A 1×10mL Substrate B 1×2mL
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 100µL Substrate Solution. Incubate 10 minutes at 37°C;
8. Read RLU value immediately.

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Magazine Citations
Microvascular Research Mutant hypoxia inducible factor-1α improves angiogenesis and tissue perfusion in ischemic rabbit skeletal muscle PubMed: 20937289
Theriogenology Evaluation of Changes in Doppler Ultrasonography Indices and Levels of Maternal Serum Angiogenic Factors throughout Pregnancy in Ewes Science: Article
Pediatr Infect Dis J Important of Angiopoietic System in Evaluation of Endothelial Damage in Children with Crimean–Congo Hemorrhagic Fever PubMed: 25831422
Oncotarget Long noncoding RNA hotair mediated angiogenesis in nasopharyngeal carcinoma by direct and indirect signaling pathways Pubmed:26717040
Experimental Animals Ginsenoside Rb1 improves cardiac function and remodeling in heart failure. pubmed:28367863
Sci Rep.  Flunarizine suppresses endothelial Angiopoietin-2 in a calcium - dependent fashion in sepsis. pubmed:28276491
PeerJ  The ratio of serum Angiopoietin-1 to Angiopoietin-2 in patients with cervical cancer is a valuable diagnostic and prognostic biomarker pubmed:28584715
Cellular Physiology and Biochemistry Angiotensin-(1-7) in Paraventricular Nucleus Contributes to the Enhanced Cardiac Sympathetic Afferent Reflex and Sympathetic Activity in Chronic Heart Failure Rats pubmed:28848201
Tumor Biology Cervical cancer cell–derived angiopoietins promote tumor progression pubmed:28720059
Anesthesiology Vasculotide, an angiopoietin-1 mimetic, restores microcirculatory perfusion and microvascular leakage and decreases fluid resuscitation requirements in … Pubmed:28968277
British Journal of Anaesthesia Vasculotide, an angiopoietin-1 mimetic, reduces pulmonary vascular leakage and preserves microcirculatory perfusion during cardiopulmonary bypass in rats 10.1016:j.bja.2018.05.049
Anticancer Research Oligodendrocytes Up-regulate the Invasive Activity of Glioblastoma Cells via the Angiopoietin-2 Signaling Pathway Pubmed: 30711932
Comparative Medicine Characterization of a Porcine Model for Von Willebrand Disease Type 1 and 3 Regarding Expression of Angiogenic Mediators in the Nonpregnant Female … Pubmed: 31526432
BMC Endocrine Disorders The role of the adipocytokines vaspin and visfatin in vascular endothelial function and insulin resistance in obese children Pubmed: 31771561
Biology of Reproduction Expression of angiogenic factors in the uteroplacental unit is altered at time of placentation in a porcine model of von Willebrand disease type 1 Pubmed: 31806575
Scientific Reports Association between Upper-airway Surgery and Ameliorative Risk Markers of endothelial function in obstructive Sleep Apnea Pubmed: 31882827
MEDIATORS OF INFLAMMATION Mica Can Alleviate TNBS-Induced Colitis in Mice by Reducing Angiotensin II and IL-17A and Increasing Angiotensin-Converting Enzyme 2, Angiotensin 1-7 … Pubmed: 33100902
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