Buffer Formulation	PBS, pH7.4, containing 0.01% SKL, 5% Trehalose.
Traits	Freeze-dried powder
Purity	> 97%
Isoelectric Point	6.5
Applications	Cell culture; Activity Assays.

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.

 

Image


Figure. Gene Sequencing (Extract)

Image


Figure. SDS-PAGE

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