大鼠骨骼肌组织雄激素受体结合容量测定方法

DETERMINATION OF BINDING ANDROGEN RECEPTOR IN RAT SKELETAL MUSCLE

目的 :以3H testosterone(T)为配基 ,测定大鼠骨骼肌胞浆中的雄激素受体结合容量。方法 :测定温度为 4℃ ,同位素配基的饱和浓度为 5 0 pmol/ml;肌组织以 4倍 (重量 /体积 )缓冲液稀释 ,0～ 4℃温度下 10 80 0 0×g离心 1h ;孵育 18～ 2 4h。结果 :骨骼肌中雄激素受体的Kd =2 ,8× 10 9mol/ml。单点法与多点法之间无显著区别。

Aim:This experiment tried to use the 3 H -testosterone (T) binding the androgen receptor (AR) in rat muscle cytosol.Methods: The temperature of binding was under 4℃, and the saturated concentration of 3 H -T was 50 pmol/ml (reaction system). The muscle tissue was homogenized with 4 vole buffer (w/v), the centrifugation of homogenate was 108 000 g and last 60 min. at 0～4℃. The time of incubation with 3 H -T was last 18～24 hours. The Kd was 2.8×10 9 mol/ml.There was not significant difference between the data of Scatchard binding plotting and single binding poltting.