The proliferation of HUVEC treated with increasing concentrations of mVEGF120 was assessed. After 72 hour treatment with mVEGF120 cells were incubated with a tetrazolium salt and the OD450-OD650 was determined.
The purity of recombinant mVEGF120 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant mVEGF120 and staining overnight with Coomassie Blue.
Western blot analysis of extracts from HUVEC untreated or treated with mVEGF120 for 15 minutes, using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 (upper) or Akt1 (C73H10) Rabbit mAb #2938 (lower)
With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg mVEGF120. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.
Stable in lyophilized state at 4°C for 1 year after receipt. Sterile stock solutions reconstituted with carrier protein are stable at 4°C for 2 months and at -20°C for 6 months. Avoid repeated freeze-thaw cycles.Maintain sterility. Storage at -20°C should be in a manual defrost freezer.
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant mVEGF120. All lots are greater than 98% pure.
Recombinant mVEGF120 contains no "tags" and the nonglycosylated protein has a calculated MW of 14,071. DTT-reduced protein migrates as a 20-26 kDa polypeptide. Lower mobility in SDS-PAGE is due to glycosylation. The non-reduced cystine-linked homodimer migrates as a 32-37 kDa protein. The expected amino-terminal APTTE of recombinant hVEGF120 was verified by amino acid sequencing.
The bioactivity of recombinant mVEGF120 was determined in a cell proliferation assay using HUVEC. The ED50 of each lot is between 1-5 ng/ml.
Less than 0.01 ng endotoxin/1μg mVEGF120.
Recombinant mouse VEGF120 (mVEGF120) Ala27-Arg146 (Accession #NP_001020428) was expressed in human 293 cells at Cell Signaling Technology.
VEGF120 , a splice variant of the VEGF-A gene, is the murine counterpart to human VEGF121 (1,2). VEGF120 is produced by endothelial cells, macrophages, T cells, and others. VEGF120 is involved in angiogenesis, vascular permeability, vascular endothelial cell survival, growth, and migration (1). VEGF120 expression is induced by hypoxia, inflammatory cytokines, and through oncogene products in tumors (1,2). VEGF120 binds to VEGFR1 and VEGFR2 receptor tyrosine kinases (1). Binding of VEGF120 to VEGFR1 and VEGFR2 leads to activation of pathways involving PI3K/Akt, p38, and FAK (1). VEGF plays a key role in tumor angiogenesis in many cancers (2).
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