Establishment of two-dimensional gel electrophoretic protein map of human spermatozoa
- VernacularTitle:人类精子蛋白质组分析的双向蛋白电泳技术研究
- Author:
Keli LUO
;
Liqing FAN
;
Guangxiu LU
- Publication Type:Journal Article
- From:
Journal of Central South University(Medical Sciences)
2001;26(2):181-184
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish the method of 2-dimensional electrophoresis(2-DE) for proteins of human spermatozoa and to construct a protein map of human spermatozoa. Methods The sperm pellet was prepared with simple Percoll layer protocol. We studied the effects of various sample preparation methods, loading quantities and isoelectric-focusing protocols on the quality of silver-stained 2-DE map, and constructed a primary protein map of human spermatozoa. Result Up to 703 protein spots were acquired with sample preparation Method Ⅰwhile only 194～210 spots with Method Ⅱ.With immobilized pH gradients and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(IPG-DALT) we could acquire over 700 spots while only 280～300 with isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(ISO-DALT). Conclusion It is satisfactory to lyse sperm with sample preparation Method Ⅰ and to separate sperm proteins by IPG-DALT for establishing 2-D map of human sperm.
