Establishment and Optimization of Two-dimensional Gel Electrophoresis Technique for Proteomics of HepG2 Cells

Objective:To optimize the treatment methods of Two-dimensional Electrophoresis sample and establish the 2-DE profiles of proteome of HepG2 cells.Methods: HepG2 cells were cultured in RPMI 1640 culture medium containing 10% fetal bovine serum,then harvested after digestion by trypsin,and lysed in lysis buffer.After centrifugation,supernatant was separated by two-dimensional gel electrophoresis,stained by Coomassie Brilliant Blue,and then followed by image scanning and analysis.Results:(297±23)protein spots were detected in the 2-DE profiles on 13 cm IPG strip(pH 3～10 L) of HepG2 cells proteome separated by 2-D PAGE.A 83% match rate of protein spots between gels was obtained.Conclusion:The establishment and optimization of two-dimensional gel electrophoresis technique for proteomics of HepG2 cells laid foundation for further research on hepatocellular carcinoma proteomics.