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目的 分离与纯化豚鼠内耳抗原 ,为寻找内耳特异性抗原打下基础。方法 采用冻融法提取豚鼠内耳抗原 ,制备性聚丙烯酰胺凝胶电泳分离与纯化内耳抗原的亚组份 ,快速染色和脱色定位蛋白区带 ,将主要蛋白区带切下匀浆回收 ,并与分析性聚丙烯酰胺凝胶电泳比较。结果 豚鼠内耳抗原的加样量为 3mg时 ,各亚组份仍能有效分离 ,三种主要亚组份 (31、4 2 - 4 5和 60kD)的相对含量分别为 2 5 99%、2 1 91%和 2 1 1%。结论 制备性PAGE可以用于纯化内耳抗原的主要亚组份。
Objective To isolate and purify the inner ear antigen of guinea pig and lay the foundation for finding the inner ear specific antigen. Methods The inner ear antigens of guinea pigs were extracted by freeze-thaw method. The sub-fractions of inner ear antigen were separated and purified by preparative polyacrylamide gel electrophoresis. The bands of proteins were rapidly stained and decolored. The major protein bands were excised and homogenized. Analytical Polyacrylamide Gel Electrophoresis Comparison. Results The results showed that the sub - components could still be effectively separated when the amount of inner ear antigen in guinea pig was 3 mg. The relative contents of the three main subunits (31, 42 - 45 and 60 kD) were 259% and 21 91% and 21.1%. Conclusions Prep PAGE can be used to purify the major sub-components of the inner ear antigen.