Active Vascular Endothelial Growth Factor Receptor 2 (VEGFR2)

CD309; FLK1; VEGFR; KDR; A Type III Receptor Tyrosine Kinase; Kinase Insert Domain Receptor; Kinase Insert Domain Receptor; Fetal Liver Kinase-1

ACTIVITY TEST

Figure. Cell proliferation of ECV-304 cells inhibit by VEGFR2.
Vascular Endothelial Growth Factor Receptor 2 (VEGFR2) also known as kinase insert domain receptor acts as a cell-surface receptor for VEGFA, VEGFC and VEGFD. VEGFR2 functions as the primary mediator of vascular endothelial growth factor activation in endothelial cells. Regulation of VEGFR-2 expression appears critical in mitogenesis, differentiation, and angiogenesis. To test the effect on inhibit the VEGF-dependent proliferation of endothelium cells, ECV-304 cells were seeded into triplicate wells of 96-well plates at a density of 5,000 cells/well and allowed to attach, replaced with serum-free overnight, then the medium was replaced with 2% serum standard DMEM including 1μg/mL Vascular Endothelial Growth Factor C (VEGFC) and various concentrations of recombinant human VEGFR2. After incubated for 96h, cells were observed by inverted microscope and cell proliferation was measured by Cell Counting Kit-8 (CCK-8). Briefly, 10µL of CCK-8 solution was added to each well of the plate, then the absorbance at 450nm was measured using a microplate reader after incubating the plate for 1-4 hours at 37℃. Proliferation of ECV-304 cells after incubation with VEGFR2 for 96h observed by inverted microscope was shown in Figure 1. Cell viability was assessed by CCK-8 (Cell Counting Kit-8) assay after incubation with recombinant VEGFR2 for 96h. The result was shown in Figure 2. It was obvious that VEGFR2 significantly inhibit cell viability of ECV-304.
(A) ECV-304 cells cultured in DMEM, stimulated with 10ng/mL VEGFR2 for 96h;
(B) Unstimulated ECV-304 cells cultured in DMEM for 96h.

Figure. VEGFR2 inhibit VEGF-dependent proliferation of ECV-304 cells.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
APMIS Peritumoral brain edema in angiomatous supratentorial meningiomas: an investigation of the vascular endothelial growth factor A pathway PubMed: 23398358
Acta Biomater Antioxidant and bone repair properties of quercetin-functionalized hydroxyapatite: an in vitro osteoblast-osteoclast-endothelial cell co-culture study PubMed: 26689470
Acta Biomaterialia Antioxidant and bone repair properties of quercetin-functionalized hydroxyapatite: An in vitro osteoblast–osteoclast–endothelial cell co-culture study Pubmed:26689470
Journal of Cellular Physiology A Human 3D In Vitro Model to Assess the Relationship Between Osteoporosis and Disseminationto Bone of Breast Cancer Tumor Cells. pubmed:27925188
Regenerative Biomaterials The directional migration and differentiation of mesenchymal stem cells toward vascular endothelial cells stimulated by biphasic calcium phosphate ceramic 10.1093/rb/rbx028
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