Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Human Vascular Endothelial Growth Factor-165 (hVEGF165 ) #8065

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Source

Recombinant human VEGF165 (hVEGF165) Ala207-Arg371 (Accession #NP_001020539) was expressed in human 293 cells at Cell Signaling Technology.

Molecular Characterization

Recombinant hVEGF165 contains no "tags" and has a calculated MW of 19,165. DTT-reduced protein migrates as a 24 kDa polypeptide and the non-reduced cystine-linked homodimer migrates as a 40 kDa protein. The expected amino-terminal APMAE of recombinant hVEGF165 was verified by amino acid sequencing.

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hVEGF165. All lots are greater than 98% pure.

Bioactivity

The bioactivity of recombinant hVEGF165 was determined in a cell proliferation assay using HUVEC. The ED50 of each lot is between 1-6 ng/ml.

Coomassie Gel

Coomassie Gel

The purity of recombinant hVEGF165 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hVEGF165 and staining overnight with Coomassie Blue.

Bioactivity

Bioactivity

The proliferation of HUVEC treated with increasing concentrations of hVEGF165 was assessed. After 72-hour treatment with hVEGF165 cells were incubated with a tetrazolium salt and the OD450 - OD650 was determined.

Western Blotting

Western Blotting

Western blot analysis of extracts from HUVEC untreated or treated with hVEGF165 for 15 minutes, using Phospho-p38 MAPK (Thr180/Tyr182) (3D7) Rabbit mAb #9215 (upper) and p38 MAPK Antibody #9212 (lower).


Endotoxin

Less than 0.01 ng endotoxin/1μg hVEGF165.

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg hVEGF165. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.

Background

VEGF165 is the most abundant splice variant of VEGF-A (1,2). VEGF165 is produced by a number of cells including endothelial cells, macrophages and T cells. VEGF165 is involved in angiogenesis, vascular endothelial cell survival, growth, migration and vascular permeability (1). VEGF gene expression is induced by hypoxia, inflammatory cytokines and oncogenes (1,2). VEGF165 binds to heparan sulfate and is retained on the cell surface and in the extracellular matrix (2,3). VEGF165 binds to the receptor tyrosine kinases, VEGFR1 and VEGFR2 (1). VEGF165 is the only splice variant that binds to co-receptors NRP-1 and NRP-2 (1-3) that function to enhance VEGFR2 signaling (1). Binding of VEGF165 to VEGFR1 and VEGFR2 leads to activation of the PI3K/AKT, p38 MAPK, FAK and paxillin (1). VEGF plays a key role in tumor angiogenesis in many cancers (2).

  1. Takahashi, H. and Shibuya, M. (2005) Clin Sci (Lond) 109, 227-41.
  2. Neufeld, G. et al. (1999) FASEB J 13, 9-22.
  3. Robinson, C.J. and Stringer, S.E. (2001) J Cell Sci 114, 853-65.

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For Research Use Only. Not For Use In Diagnostic Procedures.

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