Protein changes in human sperm demonstrated by two-dimensional electrophoresis before and after cryo

来源 :Asian Journal of Andrology | 被引量 : 0次 | 上传用户:xiaoyao984
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Objective: To establish the method of analyzing sperm protein by two-dimensional electrophoresis (2-DE) and compare the changes in protein expression before and after cryopreservation. Methods: Protein of 5×108 spermatozoa was solubilized in a 9.5 M urea lysis buffer. The sample was separated according to the isoelectric point (pI) by isoelectric focusing in the first dimension and according to the molecular weight (MW) by sodium dodecyl sulfate electrophoresis in the second dimension. The 2-D PAGE gels were stained with silver staining. Then these two protein profiles were compared. Results: The method of analyzing sperm protein by 2-DE was established and more than one protein was found to be changed after cryopreservation. Conclusion: More than one protein is changed after cryopreservation. 2-DE is an useful method to study sperm protein changes, but further improvement is needed for its practical application. Objective: To establish the method of analyzing sperm protein by two-dimensional electrophoresis (2-DE) and compare the changes in protein expression before and after cryopreservation. Methods: Protein of 5 × 108 spermatozoa was solubilized in 9.5 M urea The sample was separated according to the isoelectric focusing (pI) by isoelectric focusing in the first dimension and according to the molecular weight (MW) by sodium dodecyl sulfate electrophoresis in the second dimension. The 2-D PAGE gels were stained with Results: The method of analyzing sperm protein by 2-DE was established and more than one protein was found to be changed after cryopreservation. 2: silver staining. Then these two protein profiles were compared. -DE is an useful method to study sperm protein changes, but further improvement is needed for its practical application.
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