Establishment of the 2-D synthetic map of total protein of normal human spermatozoa enriched with low abundance protein.
- Author:
Bo-lan SUN
1
;
Li-qing FAN
;
Ling-wei LI
;
Wen-bing ZHU
;
Guang-xiu LU
Author Information
- Publication Type:Journal Article
- MeSH: Electrophoresis, Gel, Two-Dimensional; Humans; Male; Molecular Weight; Proteins; analysis; isolation & purification; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spermatozoa; chemistry
- From:National Journal of Andrology 2006;12(4):295-299
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSAll 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.
RESULTS1,080 +/- 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.
CONCLUSIONLow abundance protein was separated and a synthetic map enriched with low abundant protein was obtained by changing the protein loading amount.
