[Establishment of two-dimensional gel electrophoretic protein map of human spermatozoa].
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.


RESULTS
Up to 703 protein spots were acquired with sample preparation Method I while 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).


CONCLUSIONS
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.
Luo
K
K
Human Reproductive Engineering Laboratory, Hunan Medical College, Central South University, Changsha 410078, China.
Fan
L Q
LQ
Lu
G X
GX
D000328:Adult; D004268:DNA-Binding Proteins / Q000032:analysis*; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D008297:Male; D050809:POU Domain Factors; D013094:Spermatozoa / Q000737:chemistry*
D004268:DNA-Binding Proteins; D050809:POU Domain Factors; C484176:POU5F2 protein, human