The inducing activity of the vegetalizing factor decreases after covalent coupling to CNBr-Sepharose with reduced binding capacity. The residual inducing activity is probably due to the release of a small amount of the factor from Sepharose beads. Covalent coupling to activated CH-Sepharose completely inactivated the vegetalizing factor, whereas the neuralizing factor retained its full activity. The biological activity was also very much reduced when the vegetalizing factor was bound to Sephadex beads, a derivative of dextran. Fully active factor was recovered after enzymatic degradation of the dextran matrix with dextranase. The experiments suggest that the neuralizing factor acts on the cell surface of ectoderm cells, whereas the vegetalizing factor must probably be internalized to become biologically active.
