[Establishment of the 2-D synthetic map of total protein of normal human spermatozoa enriched with low abundance protein].
OBJECTIVE
To separate the low abundance protein and establish the 2-DE synthetic map of total protein of human normal spermatozoa by using the 2-DE technology.


METHODS
All the needed human spermatozoa were collected and mixed, and proteins were extracted at one time with the method of urea/thiourea and ultra-sound. 0.8 mg, 0.6 mg, 0.5 mg, 0.3 mg sperm protein extracts were separated with 2-DE. Analyzed with MALDI-TOF-MS, PI and MW of 2 spots were obtained. Then set the 2 spots as the referent spots, different maps were compared and analyzed. At last, a synthetic map enriched with low abundance protein was obtained.


RESULTS
1,080 +/- 23 protein spots have been separated on the 2-DE map with standard 0.5 mg loading amount and a synthetic map A was constructed which consist of 889 matched protein spots on the all maps with 0.5 mg loading amount. 381, 50 and 32 new spots were detected individually on the maps with 0.8 mg, 0.6 mg and 0.3 mg protein loading amount. A synthetic map with 1,352 protein spots was obtained.


CONCLUSIONS
Low abundance protein was separated and a synthetic map enriched with low abundant protein was obtained by changing the protein loading amount.
Sun
Bo-lan
BL
Research Institute of Reproduction and ES Cells, Central South University, Changsha, Hunan, China.
Fan
Li-qing
LQ
Li
Ling-wei
LW
Zhu
Wen-bing
WB
Lu
Guang-xiu
GX