A NEW ASSAY METHOD FOR THE DETERMINATION OF OESTROGEN RECEPTORS

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Abstract
A new assay method was developed to determine the quantity of [rabbit uterus] estrogen cytoplasmic receptors which are capable of associating with acceptor sites on chromatin. The quantity of estrogen receptor complexes associated with chromatin is of paramount importance in the mechanism of action of estrogen. This method is based on the specific binding properties of estrogen to the receptor and on the binding of the complex to chromatin. Cytosol was pre-incubated with [3H]estradiol-17.beta. ([3H]O2) to form a complex, and further incubated with a constant amount of chromatin. After this the quantity of [3]O2 cytosol receptor complexes specifically associated with the chromatin was determined. The binding activity of the chromatin to the [3H]O2 receptor complex was stable for at least 6 mo. in 0.15 M NaCl at -20.degree. C. This method enables determination of the quantity of the biologically active estrogen receptors more specifically than with other methods.