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An assay system, using the characteristic migration of follicle-stimulating hormone (FSH) on polyacrylamide gels, was developed to evaluate the effects of hypothalamic factors on amino acid incorporation into FSH in the rat adenohypophysis incubated in vitro. Follicle-stimulating hormone extracted from rat pituitary secretory granules was found to migrate on 7·5% polyacrylamide gels, pH 9·5, as a protein with an R_F of 0·614. The characteristic R_F was used to measure [14C]amino acid incorporation into FSH in rat adenohypophyses in vitro. Follicle-stimulating hormone synthesis increased after addition of crude hypothalamic extract to the incubation medium. Hormone synthesis was also increased three times after the addition of dibutyryl cyclic AMP.