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本文应用蔗糖密度梯度离心的方法进行大鼠睾丸细胞膜分离,并以此作为膜制剂进行hCG受体的检测。结果显示,用经过蔗糖密度梯度离心方法分离的睾丸细胞膜进行hCG受体放射分析,膜制剂为200μg,30℃孵育4h时,~(125)I-hCG的饱和点为4.5ng。Scatchard分析显示B_(max)=29.59fmol/mg·pr,解离常数kd=3.69×10~(-10)mol/L,具备受体可饱和、低结合容量、高亲和力的特点。因此,我们认为用蔗糖密度梯度离心的方法分离的睾丸细胞膜完全可用于hCG受体的分析。
In this paper, sucrose density gradient centrifugation method of rat testicular cell membrane separation, and as a membrane preparation for hCG receptor detection. The results showed that the hCG receptor radiosensitivity of testis membrane separated by sucrose density gradient centrifugation method was 200μg and the saturation point of ~ (125) I-hCG was 4.5ng when incubated at 30 ℃ for 4h. Scatchard analysis showed that B max = 29.59 fmol / mg · pr and dissociation constant kd = 3.69 × 10 -10 mol / L, which has the characteristics of saturation, low binding capacity and high affinity of the receptor. Therefore, we believe that testicular cell membranes isolated by sucrose density gradient centrifugation are well-suited for the analysis of hCG receptors.