Objective: To construct the two-dimensional electrophoretic (2-DE) protein map of the human sperm head.
Methods: Protein extracts of the normal human sperm and sperm head were loaded on an 18 cm immobilized pH gradients (IPG) strip holder and separated with isoelectric focusing electrophoresis as the first dimension, and then with upright SDS-PAGE as the second. Different protein spots between them were compared with Image Master 3.0, and so were the maps of method I with Urea/Thiourea/Guanidine HCl and method II with Kit/Guanidine HCl.
Results: Of the whole-sperm proteome, 802 protein spots were obtained by method I, and 797 by method II. Among them, distribution patterns of 492 spots were the same. A sperm protein map was constructed with 1107 protein spots after the interaction of the spots obtained by both the methods. Of the sperm-head proteome, 428 protein spots were obtained, all found in the whole-sperm protein map after comparison.
Conclusion: Both the methods could be complementarily used to construct a sperm protein map, and the map could serve as a model for the establishment of the sperm protein profile.