Buffer Formulation	PBS, pH7.4, containing 5% Trehalose.
Traits	Freeze-dried powder
Purity	> 90%
Isoelectric Point	9.1
Applications	Cell culture; Activity Assays.

ACTIVITY TEST

Endostatin (ES) is a naturally occurring, 20kDa C-terminal fragment derived from type XVIII collagen. It is reported to serve as an anti-angiogenic agent, similar to angiostatin and thrombospondin. Endostatin is a broad-spectrum angiogenesis inhibitor and may interfere with the pro-angiogenic action of growth factors such as basic fibroblast growth factor (bFGF/FGF-2) and vascular endothelial growth factor (VEGF). To test the effect of ES on inhibit the VEGF basic-dependent proliferation of ECV304 endothelium cell line, cells were seeded into triplicate wells of 96-well plates at a density of 5, 000 cells/well when the cell attached, replaced with serum-free standard DMEM overnight. Then the medium was replaced with 2% serum standard DMEM which contain 10ng/mL of VEGFA and various concentrations of ES. After incubated for 72h, 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 measure the absorbance at 450nm using a microplate reader after incubating the plate for 1-4 hours at 37℃. The result was shown in Figure 1. It was obvious that ES (100ng/mL) significantly inhibit cell proliferation of EV304 cells.

 

USAGE

 

Reconstitute in ddH2O to a concentration of 0.1-0.5 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. SDS-PAGE

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