The sequence of the first five N-terminal amino acids of VEGF was determined and was found to be Ala-Pro-Met-Ala-Glu.
VEGF activity was determined by the dose-dependent stimulation of the proliferation of human umbilical vein endothelial cells (HUVEC) using a concentration range of 1-8ng/ml corresponding to a Specific Activity of 125,000-1,000,000IU/mg.
The VEGF protein was lyophilized from a concentrated (1mg/ml) solution with no additives.
Sterile Filtered White lyophilized (freeze-dried) powder.
VEGF protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm using the absorbency value of 0.2875 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics).
2. Analysis by RP-HPLC, using a calibrated solution of VEGF as a Reference Standard.
Greater than 98.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Analysis by SDS-PAGE.
It is recommended to reconstitute the lyophilized VEGF in sterile 18MΩ-cm H2O not less than 100ug/ml, which can then be further diluted to other aqueous solutions.
Lyophilized VEGF although stable at room temperature for 3 weeks, should be stored desiccated below
-18oC. Upon reconstitution VEGF should be stored at 4oC between 2-7 days and for future use below -18oC.
For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please prevent freeze-thaw cycles.
Vascular endothelial growth factor A, VEGF-A, Vascular permeability factor, VPF, VEGF, MGC70609.
