Isolation and purification of guinea pig inner ear antigens by preparative polyacrylamide gel electrophoresis

制备性聚丙烯酰胺凝胶电泳分离与纯化豚鼠内耳抗原(英文)

Objective To isolate and purify the subcomponents of guinea pig inner ear antigens for further study on the autoimmunity of the inner ear.Methods Inner ear tissues were homogenized in phosphate buffered saline containing 0.1% SDS, and then frozen and defrosted repeatedly to extract inner ear antigens. Preparative polyacrylamide gel electrophoresis was used to separate the subcomponents of inner ear antigens. Following electrophoresis,the protein bands were localized by rapid staining and destaining.Results The major protein bands were clearly distinct when 3 mg of crude inner ear antigens was loaded,and the three major subcomponents (31, 42- 45 and 60 kD proteins) accounted for about 25.99%,21.91% and 21.10%, respectively.Conclusion Preparative polyacrylamide gel electrophoresis can be used to purify the major subcomponents of inner ear antigens.

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。