论文部分内容阅读
分离的微粒体已在生化和药理研究中得到广泛应用。为了使不同实验室或同一实验室于不同时间的实验结果可相互比较,应有标准化的制剂。然而,由于用同一方法得到高纯度、高得率的微粒体很为困难,这一问题可通过保存已知纯度及活性的微粒体制剂来解决。本文探讨了保存微粒体酶活性的各种条件。选用220±10g 大鼠,取肝脏制成匀浆,加入50mMpH7.4的磷酸缓冲液(内含154mM 的氯化钾)高速离心,取9000×g 部位(4ml/g 组织)。将此部作于10500×g 离心30分钟或钙沉淀法制备微粒体。实验结果表明,不同分离条件对微粒体酶活性的影响很小。新鲜肝脏在0°保存24、48、72小时后再制备9000×g 部位,对苯胺的羟化、4-硝基苯甲醚的0-脱甲
Isolated microsomes have been widely used in biochemical and pharmacological studies. In order to make different laboratories or laboratories at different times the experimental results can be compared with each other, there should be standardized formulations. However, since it is very difficult to obtain high-purity, high-yield microsomes by the same method, this problem can be solved by keeping microsome preparations of known purity and activity. This article explores various conditions for preserving microsomal enzyme activity. 220 ± 10g rats were selected and the liver was homogenized and centrifuged at a speed of 9000 × g (4ml / g tissue) by adding 50mMpH7.4 phosphate buffer (containing 154mM potassium chloride). This section was made at 10500 × g centrifugation for 30 minutes or calcium precipitation method to prepare microsomes. The experimental results show that different separation conditions have little effect on microsomal enzyme activity. The fresh liver was stored at 0 ° for 24, 48, and 72 hours prior to preparation of a 9000 × g site for hydroxylation of aniline and 0-demethylation of 4-nitroanisole