【摘要】 目的:对细胞膜色谱固定相的制备条件进行了优化,在此基础上提出了标准操作流程。方法:以兔红细胞为原材料,使用超声破碎法获取细胞膜,减压震荡法进行细胞膜固定化,通过BCA法测定细胞膜蛋白含量,研究细胞膜加入量、真空度、静置时间和载体材料对细胞膜色谱法的影响。结果:确定制备细胞膜色谱固定相的最优条件是细胞膜蛋白加入量为80μg(膜蛋白)/mg(硅胶),真空度为0.075 MPa,静置时间为14 h,并采用多孔硅胶。结论:通过试验考查出了细胞膜色谱固定相制备的最优条件和标准操作流程。更多还原

【Abstract】 Objective: To get a standard protocol for CMSP by optimizing the preparation conditions. Method: Rabbit red cell membranes were prepared by a sonication method and immobilized by a reduced-pressure method. A BCA method was adopted for determination of membrane proteins immobilized onto the CMSP. Effects of amount of cell membranes,applied vacuum,standing time as well as carrier materials on the preparation of CMSP were systematically investigated. Results: Optimum conditions for preparation of CMSP include 80 μg( membrane proteins)/mg( silica beads),0. 075 Mpa of the applied vacuum,14 h of the standing time,and porous silica beads used for CMSP preparation. Conclusions: In this work,standard operation conditions and protocols for the preparation of CMSP were provided,after careful investigation of the effects of various factors on CMSP’s preparation.更多还原