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

Objective To isolate and purify the subcomponents of guinea pig inner ear antigens for fur ther 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. Pr eparative polyacrylamide gel electrophoresis was used to separate the subcompone nts of inner ear antigens. Following electrophoresis, the protein bands were lo calized by rapid staining and destaining. Results The major protein bands were clearly distinct when 3 mg of crude inner ear anti gens was loaded, and the three major subcomponents (31, 42-45 and 60 kD protein s) accounted for about 25. 99%, 21. 91% and 21. 10%, respectively. Conclusion Preparative polyacrylamide gel electrophoresis can be used to purify the major s ubcomponents of inner ear antigens.