人心肌肌钙蛋白T的提取和纯化及初步鉴定

Purification and Identification of human Myocardial Troponin T

目的 从人心肌组织中提取、纯化心肌肌钙蛋白T ,并进行初步鉴定。方法 采用匀浆、高盐提取、柱层析的方法 ,从心肌组织中分离、纯化cTnT ,以凝胶电泳、免疫印迹进行鉴定。结果 cTn粗品经柱层析纯化 ,其第Ⅱ峰为cTnT活性峰 ,分子量约为3 4kD。结论 本实验获得纯度较高的cTnT ,为进一步实验研究奠定了基础。

Objective To obtain high pure myocardiac Troponin T from myocardiac tissue. Methods After the myocardiac tissues were homogenized and purified with high salt solution, the Sephacryl S-300HR column chromatography was employed to separate the cTnT from cardiac proteins, and identified them with SDS-PAGE and Western blot. Results The crude cTn was purified by column chromatography, and there appeared three peaks. The second peak was single component, which could react with mouse anti-human Troponin T antibody, and its molecular weight was about 34kD. Conclusions In this experiment, high pure cTnT was obtained, which could pave a way for further research.