Multiplex Assay Kit for Vascular Endothelial Growth Factor C (VEGFC) ,etc. by FLIA (Flow Luminescence Immunoassay)

VEGF-C; Flt4-L; VRPL; Vascular Endothelial Growth Factor-Related Protein

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Vascular Endothelial Growth Factor C (VEGFC) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Vascular Endothelial Growth Factor C (VEGFC) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Vascular Endothelial Growth Factor C (VEGFC) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Vascular Endothelial Growth Factor C (VEGFC) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 93-102 97
EDTA plasma(n=5) 86-101 90
heparin plasma(n=5) 95-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Vascular Endothelial Growth Factor C (VEGFC) ,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 Vascular Endothelial Growth Factor C (VEGFC) ,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 Vascular Endothelial Growth Factor C (VEGFC) ,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) 89-104% 81-97% 84-97% 92-105%
EDTA plasma(n=5) 78-92% 95-102% 87-95% 92-101%
heparin plasma(n=5) 98-105% 89-98% 93-101% 86-95%

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#:VEGFC) 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
Journal of Cellular Biochemistry Alternatively activated RAW264. 7 macrophages enhance tumor lymphangiogenesis in mouse lung adenocarcinoma PubMed: 19241443
genetics and molecular research Expression of COX-2 and VEGF-C in cholangiocarcinomas at different clinical and pathological stages PubMed: 26125824
Experimental and Therapeutic Fenofibrate inhibits the expression of VEGFC and VEGFR-3 in retinal pigmental epithelial cells exposed to hypoxia PubMed: 26622498
Am J Pathol IL-10 Indirectly Regulates Corneal Lymphangiogenesis and Resolution of Inflammation via Macrophages PubMed: 26608451
PLoS One. Interleukin-6 Induces Vascular Endothelial Growth Factor-C Expression via Src-FAK-STAT3 Signaling in Lymphatic Endothelial Cells. pubmed:27383632
Oncology Letters RNAi-mediated gene silencing of vascular endothelial growth factor C suppresses growth andinduces apoptosis in mouse breast cancer in vitro and in vivo. pubmed:27895746
Angiogenesis Characterization of isolated liver sinusoidal endothelial cells for liver bioengineering Pubmed:29582235
Oncology Reports Effects of diphyllin as a novel V-ATPase inhibitor on TE-1 and ECA-109 cells Pubmed:29328465
LABORATORY INVESTIGATION Dynamic signature of lymphangiogenesis during acute kidney injury and chronic kidney disease Pubmed: 31019289
Cutaneous and Ocular Toxicology Isotretinoin does not alter VEGF-A and VEGF-C levels: Do retinoids behave differently in dose-dependent and/or in vivo/in vitro conditions? Pubmed: 32722957
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