Vascular endothelial growth factor (VEGF) is homodimeric, heparin-binding glycoprotein involved in both angiogenesis and vasculogenesis. VEGF is expressed as multiple alternately spliced isoforms of VEGF121, 165, 189 and 206. VEGF binds to the receptor tyrosine kinases VEGF R1 (Flt-1) and VEGF R2 (KDR/Flk-1) to activate signal transduction and regulate both physiological and pathological angiogenesis. Source: Recombinant protein corresponding to aa207-327 from human VEGF121, expressed in E. coli. Molecular Weight: ~14.2kD (122aa) Biological Activity: Measured in a cell proliferation assay using NIH-3T3 cell, the ED50 for this effect is <1.8ng/ml. Activity Assay: 1. Cell Line: NIH-3T3 (Mus mosculus/ embryo) 2. Maintenance Condition: RPMI 1640 containing 10% FBS 3. Assay Medium: serum free RPMI 1640 4. Cell density: 2x10e4 cells/well (96 well plate, final volume 100ul) 5. Serum Free Starvation: 24hr with RPMI1640 6. Incubation Time: 24 hr (after sample treatment) 7. Concentration Range: 100pg/ml-10ug/ml 8. Detection method: BrdU assay Amino Acid Sequence: MAPMAEGGGQ NHHEVVKFMD VYQRSYCHPI ETLVDIFQEY PDEIEYIFKP SCVPLMRCGG CCNDEGLECV PTEESNITMQ IMRIKPHQGQ HIGEMSFLQH NKCECRPKKD RARQEKCDKP RR Endotoxin: ~1EU/1ug of protein (LAL) Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 6 months after receipt at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
