[Study on the method of two dimensional polycrylamide gel electrophoresis on rat condylar chondrocyte].
OBJECTIVE
To investigate the protein profile by two dimensional polycrylamide gel electrophoresis on the rat condylar chondrocyte in vitro.


METHODS
The third-passage chondrocytes were harvested from the mandibular condyles of 2-day-old rats in this study. The protein profile of the rat mandibular condylar chondrocytes was examined by two dimensional polycrylamide gel electrophoresis (2-DE-PAGE). The 2-DE gel maps on different pH gradients were obtained. The result of modified coomassi blue-sliver staining and sliver staining was compared using Pdquest 7.1 image analysis software.


RESULTS
The results showed that the good protein profile of the condylar chondrocytes was obtained by standard Bio-Rad manual. The protein was mainly in the field from pH4 to pH7. The 1203±86 protein points were examined on 2-DE gel map by modified coomassi blue-sliver staining, and 1769±97 protein points was examined by sliver staining. The silver staining map showed more distinctly but higher background than modified coomassi blue-sliver staining.


CONCLUSIONS
The protein profile of the condylar chondrocytes enriches the proteomic database and gives evidence to further proteomic research. The 2-DE map obtained by modified coomassi blue-sliver staining is more suitable for MALDI-TOF mass identification. Supported by National Natural Science Foundation of China (Grant No. C30700963), China Postdoctoral Science Foundation(Grant No.20090461088), Jiangsu Provincial Postdoctoral Science Foundation (Grant No.0802003C) and Nanjing City's Science and Technology Foundation (Grant No.200905011).
Wu
Tuo-jiang
TJ
Department of Orthodontics, the Affiliated Stomatological Hospital of Nanjing Medical University, Nanjing 210029, China. wutuojiang@163.com
Li
Huang
H
Ma
Qiao-lin
QL
Wang
Wen-mei
WM
D000818:Animals; D019902:Chondrocytes; D015180:Electrophoresis, Gel, Two-Dimensional; D008335:Mandibular Condyle; D040901:Proteomics; D051381:Rats