RecombinantVEGF164,Mouse(P.pastoris-expressed)

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.