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作者介绍的方法具有快速、简易、重复性好等特点。将6μ厚未固定的冰冻组织切片置于用明胶——铬矾包被的盖玻片上,置室温干燥后,用100μlTED缓冲液(含0.1～8.0nM[~3H]-雌二醇,测非特异性结合时再加200倍过量的己烯雌酚)复盖在切片上。分别于4℃、23℃及37℃保温1～20小时。保温结束后,组织切片用TED缓冲液冲洗,再用液闪法直接测定切片结合的受体量。切片同时作组织学染色,并按Lowvy法测定蛋白质含量。Scatchard分析表明,靶组织(牛及大鼠子宫)的冰冻切片的复盖液和冲洗后的切片中均存在高亲和力、低容量的Ⅰ型雌激素结合位点,解离常数为
The method introduced by the author has the characteristics of quickness, simplicity and repeatability. Six micrograms of unfixed frozen tissue sections were placed on coverslips coated with gelatin-chrome alum and left to dry at room temperature. After 100 μl of TED buffer (0.1 ~ 8.0 nM [~ 3H] -estradiol) Heterosexual combination plus 200 times excess diethylstilbestrol) covered in the slice. Respectively, at 4 ℃, 23 ℃ and 37 ℃ for 1 to 20 hours. After incubation, tissue sections were washed with TED buffer and the amount of bound receptor bound was measured directly by liquid scintillation. Slices were also histologically stained and the protein content determined by the Lowvy method. Scatchard analysis showed that both the high affinity and low capacity type I estrogen binding sites existed in the frozen sections of the target tissues (bovine and rat uterus), and the dissociation constants were