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  • Title: The measurement of cytosolic estrogen receptors in human endometrial tissue and organ cultures.
    Author: Rodgers NT, Kaufman DG.
    Journal: J Steroid Biochem; 1981 Aug; 14(8):801-6. PubMed ID: 7300349.
    Abstract:
    This paper describes a method for determining the number of cytosolic estrogen receptors in small quantities of human endometrial tissue. The method is a modification of Puca et al.'s gel filtration technique developed for measurement of estrogen receptors in whole calf uteri. Sucrose gradient ultracentrifugation was used to determine sedimentation coefficients in the estrogen binding proteins of endometrial cytosol. Scatchard and double reciprocal analyses were used to determine graphically the total number of binding sites and dissociation constants. Estrogen receptor levels were measured in 22 patients at various points in the menstrual cycle. The highest receptor levels among the patients were observed in the endometrium histologically determined to be in the middle of the proliferative phase of the menstrual cycle. The quantity of the receptors decreased dramatically during the last part of the proliferative phase, the reduced levels persisting during the secretory phase. Estrogen receptors were also found in organ cultures of human endometrium maintained in vitro for up to 67 days. Estrogen receptors were found to have a sedimentation coefficient of approximately 8S, a dissociation constant of approximately 1 x 10 -9 molar, and a binding capacity ranging from 2.3 x 10 -13 to 8 x 10 -15 mol/mg of protein. This study shows that receptor assays can be performed with 100-200 mg of tissues using 10 mM triated hydrogen-estradiol binding and gel filtration method. The assay reported here may also be useful in measuring estrogen receptors of small clinical specimens, and in analyzing needle biopsy specimens or currettings.
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