[Analysis of the difference in NIH3T3 cell protein expression profiles before and after TPA treatment using two-dimensional polyacrylamide gel electrophoresis and image analysis software].
OBJECTIVE
To establish and optimize two-dimensional (2-D) polyacrylamide gel electrophoresis (PAGE) for comparative analysis of the protein expression profiles in mouse fibroblast NIH3T3 cells before and after 12-O-tetradecanoy- lphorbol-13-acetate (TPA) treatment using image analysis software, so as to prepare for more intensive study in the identification of the proteins that mediate the biological effect of TPA.


METHODS
The total cellular protein extracted from NIH3T3 cells with or without TPA treatment underwent 2-D PAGE and silver nitrate staining prior to the analysis of the differential protein expressions using image analysis software.


RESULTS
Image analysis revealed obvious differential protein expressions of the cells in response to TPA treatment.


CONCLUSIONS
High-quality instruments for 2-D PAGE, skilled electrophoretic operation and efficient image processing are all essential in the reliable identification of stable functional proteins.
Shen
Xin-ming
XM
Institute of Cancer Research, First Military Medical University, Guangzhou 510515, China.
Jiang
Pei-zhou
PZ
Huang
Hua
H
Li
Xin
X
Yao
Kai-tai
KT
D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D007091:Image Processing, Computer-Assisted; D051379:Mice; D041681:NIH 3T3 Cells; D011506:Proteins / Q000032:analysis*; D012984:Software; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
D011506:Proteins; D013755:Tetradecanoylphorbol Acetate