| A Method for the Determination of Estrogen Receptor Level in Frozen Sections of Porcine Uterus |
|
Yong-Dal Yoon1, Chor-Hong Park1, Young-Keun Lee2 |
1Department of Biology, College of Natural Sciences , Hanyang University
2Radiation Application Department, Korea Advanced Energy Research institute |
| 냉동절편을 이용한 돼지 자궁내 에스트로겐 수용체의 측정 |
|
윤용달1, 박철홍1, 이영근2 |
1한양대학교 자연과학대학 생물학과
2한국에너지연구소 방사션응용연구실 |
|
|
| Abstract |
|
The present study was designed to develop a new method for the determination of estrogen receptor in porcine uterus using frozen sections. Cryostat sections were incubated with $^3H$-estradiol($^3H$-$E_2$) in the presence or absence of diethylstilbestrol(DES) and the radioactivity of 3H-E2 bound to estrogen receptor(ER) was detected. The level of specific estrogen receptor was determined by Scatchard analysis. The highest ratio of specific binding against total binding was achieved in 3 sec. tions(5mm x 5mm) which was corresponded to lOO${\mu}$/ml protein concentration. Optimal binding was obtained during incubation with $^3H$-$E_2$ for 30 minutes at 23$^{\circ}C$ after treatment of sections with acetone for 20 seconds. Three time-washing of sections was proved to be appropriate for the removal of unbound 3H-E2. 200-fold molar excess of DES was substituted for the binding of $^3H$-$E_2$ to ER sufficiently(binding efficiency of 54.8%). ER was saturated with 4nM of $^3H$-$E_2$ and its dissociation constant was 0.1nM. ER assay using frozen sections(Histological radioreceptor assay, HRRA) was significantly correlated with radioreceptor assay for estradiol(RRA, 0.976 , p
|
|
PDF Links
Full text via DOI
Download Citation
