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在完整细胞与3H-R1881特异结合的基础上,用STM-TritonX-100缓冲液分离粗制核,建立了简便而快速地检测核雄激素特异结合的方法。对正常男性包皮成纤维细胞核与3H-R1881特异结合的Scatchard及单点分析表明,该细胞核与3H-R1881的特异结合具有高亲和力、低容量的特征,当细胞与近饱和浓度的3H-R1881温育50min后,47.16±7.99%(n=20)的特异结合被发现存在于核内,而一睾丸女性化(tfm)综合征患者的核特异结合量明显低于该值。我们还显示,正常男性包皮成纤维细胞与3H—R1881在42℃温育时的特异结合量比在37℃温育时减少,但其量低于40%。当减少量超过40%时,表明存在热不稳定型的雄激素受体。
Based on the specific binding of intact cells to 3H-R1881, the crude nuclei were separated by STM-TritonX-100 buffer, and a simple and rapid method for detecting the specific binding of nucleoside and androgen was established. Scatchard and single point analysis of the specific binding of normal male foreskin fibroblast nuclei to 3H-R1881 showed that the specific binding of 3H-R1881 to this nucleus was characterized by high affinity and low volume. When the cells were incubated with 3H-R1881 near-saturated concentration After 50 min incubation, specific binding of 47.16 ± 7.99% (n = 20) was found to be present in the nucleus, whereas the nuclear specific binding of t testosterone syndrome was significantly lower in this testicle. We also show that the specific binding capacity of normal male foreskin fibroblasts to 3H-R1881 when incubated at 42 ° C is less than when incubated at 37 ° C but less than 40%. A decrease of more than 40% indicates the presence of heat-labile androgen receptor.