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. CONCLUSION: 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.