Resuspend cells in assay media (Earle Salts 199X Gibco #11150, without h-EGF) containing 10 % HI-FBS and transfer 100 ul/well to assay plate (5,100 cells/well passage#2). Serial dilute murine VEGF in assay media containing 10 % HI-FBS and transfer 100ml/well to cells in assay plate. Final assay volume is 200 ml/well, containing 10 % HI-FBS, and murine VEGF as indicated 89 hours. Add 20 ml of Promega Substrate Cell Titer 96 Aqueous One Solution Reagent to each well, Incubate 37°C and read at OD 490 nm. The ED50 for the effect is typically 4.1 - 6.2 ng/ml.
Pureté
> 97 % by SDS-PAGE.
niveau d'endotoxine
level is less than 0.1 ng per myg (1 EU/myg) of VEGF
VEGF is a potent growth and angiogenic cytokine. It stimulates proliferation and survival of endothelial cells, and promotes angiogenesis and vascular permeability. Expressed in vascularized tissues, VEGF plays a prominent role in normal and pathological angiogenesis. Substantial evidence implicates VEGF in the induction of tumor metastasis and intra-ocular neovascular syndromes. VEGF signals through the three receptors, fms-like tyrosine kinase (flt-1), KDR gene product (the murine homolog of KDR is the flk-1 gene product) and the flt4 gene product. Synonym: VEGF 6. Formulation: Lyophilized from 25 mM HEPES and 150 mM NaCl, pH 7.0.