Human vitronectin (478aa) is a liver derived major plasma glycoprotein that exhibits multiple activities and functions as a cell adhesion molecule and regulator of coagulation. It contains the amino acid structural motif, Arg-Gly-Asp (RGD), which is involved in cell attachment. Human Vitronectin has a mass of 75kDa and circulates as a single-chain moiety of 75kDa and a two-chain moiety of 65kDa and 10kDa. Vitronectin belongs to the group of structurally and functionally homologous adhesive proteins (fibrinogen, fibronectin, Von Willebrand factor), which interact with platelets and the vessel wall in the early stages of blood clotting. When coated on surfaces, very low concentrations of Vitronectin promote endothelial cell attachment and induce spreading and migration of cells in a time- and concentration-dependent fashion. Recent data from James Thomson’s laboratory indicated that deletion of C terminal of vitronectin domain displays its capacity as matrix coating protein for both ES and iPS cell cultures when combined with E8 medium(1).

N-terminal human Vitronectin gene (62 - 398 aa) was constructed with codon optimization and expressed in non-fusion protein form in E.coli as inclusion bodies. The final product was refolded using our unique “temperature shift inclusion body refolding” technology and chromatographically purified. Coating this recombinant protein E8, ES or NutriStem medium at concentration of 0.5 ug – 1 ug per cm2 will benefit for cultures of either human ES cell or iPS cells.

1. Purity: > 95% by SDS-PAGE.

2. Biological Activity: When coated onto tissue culture plastics, Human Vitronectin promotes one half maximal attachment of BALB/3T3 cells in serum-free medium at < 0.1µg / cm2. Maximum attachment should occur at approximately 0.2µg / cm2.

Each Lot was tested using 10ug/well (in 6 well plate) in Nutristem medium for human ES cells (H9).