Thesis

Electrophoretic extraction of adhesion-inhibiting protein

An aggregation inhibitory factor (AIF) has been extracted from mouse ascites teratoma cells that retards adhesion of cultured teratoma cells of the same cell line. Preliminary characterization of AIF on polyacrylamide gels suggests that AIF is a carbohydrate-free protein composed of four subunits. Extraction of AIF from ascites teratoma cells was accomplished without significant loss of viability by a novel technique involving the application of an electric field to large numbers of whole cells suspended in a hypertonic electrode buffer. Assumptions were made regarding the force developed by this field on charged transmembrane proteins in order to measure the potential of this technique for extracting proteins of possible significance in adhesion. In tests of adhesion, AIF consistently inhibited auto-aggregation of cultured teratoma cells after 5, 10, 15, and 30 minutes of incubation with cells. Furthermore, a reduced concentration of AIF resulted in a corresponding decrease in inhibition, suggesting a concentration dependent action. Further work with AIF may reveal if this factor plays a role in the adhesion mechanism of cultured teratoma cells.

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