技术与方法 |
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The study on separating the protein of bovine sperm by two-dimensional gel electrophoresis |
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Abstract Two-dimensional gel electrophoresis is a protein separation technique and it can be used for separating sperm proteins. The objectives of this study were to develop an optimal two-dimensional gel electrophoresis, and the bovine sperm proteins were isolated with the developed technique. In this study, isoelectric focusing electrophoresis program was optimized, and various sample preparation methods, different loading quantities and strip length were studied. The results indicated that the better proteins map was obtained when the bovine sperm were lysed by two step method of urea-guanidine hydrochloride and the 13cm N-linear IPG strip was used to conduct the isoelectric focusing electrophoresis. More than 800 protein spots could be detected by the imaging analysis system, and the analysis result indicated that molecular weight and isoelectric point of most proteins were from 10 to 100 KD and 4.0 to 9.0 respectively. The study provided a basis for detecting and analyzing the differential proteins of the X,Y sperm.
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Received: 11 April 2006
Published: 25 September 2006
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