Multiplex Assay Kit for Angiopoietin 1 (ANGPT1) ,etc. by FLIA (Flow Luminescence Immunoassay)

ANG1; AGP1; AGPT; ANG-I

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Angiopoietin 1 (ANGPT1) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Angiopoietin 1 (ANGPT1) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Angiopoietin 1 (ANGPT1) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Angiopoietin 1 (ANGPT1) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 97-105 101
EDTA plasma(n=5) 86-103 98
heparin plasma(n=5) 82-96 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Angiopoietin 1 (ANGPT1) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Angiopoietin 1 (ANGPT1) ,etc. by FLIA (Flow Luminescence Immunoassay) 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 1 (ANGPT1) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 84-91% 93-101% 99-105% 93-104%
EDTA plasma(n=5) 86-99% 88-102% 86-101% 78-102%
heparin plasma(n=5) 87-97% 84-91% 78-89% 79-98%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:ANGPT1) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Theriogenology Evaluation of Changes in Doppler Ultrasonography Indices and Levels of Maternal Serum Angiogenic Factors throughout Pregnancy in Ewes Science: Article
J Atheroscler Thromb High Mobility Group Box 1 Promotes Angiogenesis from Bone Marrow-derived Endothelial Progenitor Cells after Myocardial Infarction PubMed: 25735431
Archives of Biological Sciences Diabetes-induced renal failure is associated with tissue inflammation and neutrophil gelatinase-associated lipocalin: Effects of resveratrol publication:299492265_Diabetes-induced_renal_failu
Am J Transl Res Angiopoietin-1 facilitates recovery of hematopoiesis in radiated mice Pubmed:27347310
Clinical research Increased plasma concentrations of interleukin 35 in patients with coronary artery disease Journal:-19
Turkish Journal of Medicl Sciences A new angiogenesis prognostic index with VEGFA, PlGF, and angiopoietin 1 predicts survival in patients with advanced gastric cancer pubmed:28425270
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
Molecular&cellular proteomics Prediction of Response to Sorafenib in Hepatocellular Carcinoma: A Marker Panel by Multiple Reaction Monitoring-Mass Spectrometry pubmed:28550167
Tumor Biology Cervical cancer cell–derived angiopoietins promote tumor progression pubmed:28720059
Free Patents online RHEUMATOID ARTHRITIS RELATED BIOMARKER 20170350884
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
The Journal of Biological Chemistry Toll-like receptor 2 deficiency hyperactivates the FoxO1 transcription factor and induces aging-associated cardiac dysfunction in mice Pubmed:29929978
Clinical Significance of Nectin-4 Expression in Metastasis and Angiogenesis for Tumor Relapse Doi: 10.4143/crt.2018.214
BIOLOGY OF REPRODUCTION Prostaglandin-mediated effects in early canine corpus luteum: In vivo effects on vascular and immune factors Pubmed: 30929911
Scientific Reports Association between Upper-airway Surgery and Ameliorative Risk Markers of endothelial function in obstructive Sleep Apnea Pubmed: 31882827
Biochem Biophys Res Commun Effects of interleukin-6 and tumor necrosis factor-α on the expression of angiogenic and collagenolytic factors in premature and mature adipocytes Pubmed: 32800538
J Nutr Biochem Effects of protein malnutrition on haematopoietic regulatory activity of bone marrow mesenchymal stem cells 33705953
Intensive Care Med Exp The effect of targeting Tie2 on hemorrhagic shock-induced renal perfusion disturbances in rats 33997943
Gynecol Endocrinol Potential of thrombospondin-1 in treatment of polycystic ovary syndrome rat model: a preliminary study 34282706
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RPA008Hu01 Recombinant Angiopoietin 1 (ANGPT1) Positive Control; Immunogen; SDS-PAGE; WB.
RPA008Hu02 Recombinant Angiopoietin 1 (ANGPT1) Positive Control; Immunogen; SDS-PAGE; WB.
RPA008Hu03 Recombinant Angiopoietin 1 (ANGPT1) Positive Control; Immunogen; SDS-PAGE; WB.
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MAA008Hu21 Monoclonal Antibody to Angiopoietin 1 (ANGPT1) WB
MAA008Hu22 Monoclonal Antibody to Angiopoietin 1 (ANGPT1) WB; IHC; ICC; IP.
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