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Korean Journal of Fertility and Sterility 1986;13(2):187-193.
Determination of Potimal Conditions for Cytosolic Progesterone Receptor Assay in Rat Uterus
TK Park1, KH Park1, K Seo1, KS Rhee2, MC Yun2, KJ Ryu2
1Department of Obstetrics and Gynecology, Yonsei University College of Medicine
2Department of Pharmacology, Yonsei University College of Medicine
흰쥐 자궁에서의 Cytosolic Progesterone 수용체 측정에 관한 연구
박찬규1, 박기현1, 서경1, 이건수2, 윤미정2, 유경자2
1연세대학교 의과대학 산부인과학교실
2연세대학교 의과대학 약리학교실
Abstract
The purpose of this study was to establish optimal conditions for progesterone receptor assay using rat uterus, therby applying this system to understand action mechanism of progesterone in female reproductive organ and to evaluate physiological and pathological conditions in female reproduction. The results obtained were as follows 1. $^3H-R5020$ seemed to be more stable than 3H-progesterone as a ligand. 2. Optimal incubation time for ligand and receptor binding was 4-5 hrs at $4^{\circ}C$. 3. For the separation of bond and free ligand, optimal charcoal incubation time was 20 mins. 4. 2-3 mg cytosolic protein/ml was optimal for the binding of ligand. 5. Endogenous progesterone did not influence binding of ligand and receptor unless endogenous progesterone levels were extremely high as in case of pregnancy. 6. Dissociation rate for progesterone receptor was 1.22 nM. 7. $^3H-R5020$ did not bind to corticoid binding globulin (CBG), suggesting that addition of cortisol is saturate CBG was, not necessary as far as $^3H-R5020$ was used as a radioligand. It is, therefore, considered that this assay system established with these conditions might be used for the research as well as clinical routine purposes.
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