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ELISA Kit for Vascular Endothelial Growth Factor A (VEGFA)

VEGF; VEGF-A; VPF; Vascular Endothelial Cell Growth Factor A; Vascular Permeability Factor

Specificity

This assay has high sensitivity and excellent specificity for detection of Vascular Endothelial Growth Factor A (VEGFA).
No significant cross-reactivity or interference between Vascular Endothelial Growth Factor A (VEGFA) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Vascular Endothelial Growth Factor A (VEGFA) and the recovery rates were calculated by comparing the measured value to the expected amount of Vascular Endothelial Growth Factor A (VEGFA) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 97-105 102
EDTA plasma(n=5) 86-101 93
heparin plasma(n=5) 96-103 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Vascular Endothelial Growth Factor A (VEGFA) 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 A (VEGFA) 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 A (VEGFA) 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) 83-98% 89-96% 79-91% 95-104%
EDTA plasma(n=5) 90-104% 80-99% 96-103% 85-99%
heparin plasma(n=5) 86-98% 86-101% 85-101% 80-95%

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
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 Reference
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Acta Biomaterialia Multiple release of polyplexes of plasmids VEGF and bFGF from electrospun fibrous scaffolds towards regeneration of mature blood vessels PubMed: 22484697
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Nanoscale Research Letters Silver nanoparticles administered to chicken affect VEGFA and FGF2 gene expression in breast muscle and heart. PubMed: 22827927
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PLoS One. Molecular Effects of Doxycycline Treatment on Pterygium as Revealed by Massive Transcriptome Sequencing PubMed: PMC3378547
Oncology Reports A bone-seeking clone exhibits different biological properties from the ACHN parental human renal cell carcinoma in vivo and in vitro PubMed: PMC3583590
The Journal of Biological Chemistry Intertissue Flow of Glutathione (GSH) as a Tumor Growth-promoting Mechanism: INTERLEUKIN 6 INDUCES GSH RELEASE FROM HEPATOCYTES IN METASTATIC B16 MELANOMA-BEARING MICE PubMed: PMC3091180
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Stem Cells International Matrix Metalloproteinase 9 Secreted by Hypoxia Cardiac Fibroblasts Triggers Cardiac Stem Cell Migration In Vitro Pubmed:25767513
Physiology and Pharmacology A Study on the Serum Levels of Angiogenic Factors in Response to Acute Long-term Submaximal Exercise in Sedentary Men Phypha:Source
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American Journal of Biomedical Sciences Insulin Like Growth Factor -1(IGF-1) Promotes Angiogenesis and Reverses Ischemia Reperfusion Induced Acute Kidney Injury in Rats: Role of VEGF and TGF-β1 publication:303524932_Insulin_Like_Growth_Factor_-
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