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Published on: January 20, 2019
Isolation and purification of guinea pig inner ear antigens by preparative polyacrylamide gel electrophoresis
1Department of Otolaryngology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. keyc@tjmu.edu.cn
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.
