Abstract
Two-dimensional (2D) gel electrophoresis is a powerful technique used for the differential protein expression analysis. High-resolution two-dimensional electrophoresis is still the main applied separation technique in proteomics. A successful 2D gel electrophoresis depends on the proper optimized conditions most importantly an optimized buffer. In this study we checked different factors affecting 2D gel results including rehydration time, reducing agents, chaotrophic agents, and detergents. We chose Cynodon dactylon as an experimental plant. Proteins were extracted and purified. For the proper solubilization of the protein sample different rehydration buffers of pH 3-10 were applied having different amount of chaotrophic agents like urea and thiourea, detergents (NP-40, Triton X-100 and CHAPS) and reducing agents (β-merceptoethanol and DTT). To further improve the protein profile on 2D gel, solubilized proteins were re-extracted, washed and re-solubilized in a fresh rehydration buffer. Our modified protocol showed a high increase in protein solubility and improvement in protein 2D gel profile for C. dactylon proteome. In conclusion, we have successfully optimized the conditions for C. dactylon proteome analysis by 2D gel electrophoresis. These optimized conditions will lay the basis for further profound studies in the field of proteomics.
Highlights
Concurrent chemoradiotherapy (CRT) has been reported to significantly increase the survival rate of patients with esophageal cancer as compared with radiation therapy alone [1,2]
One hundred and fourteen patients were treated with CRT
Performance status and body weight loss were identified as significant prognostic factors
Summary
Concurrent chemoradiotherapy (CRT) has been reported to significantly increase the survival rate of patients with esophageal cancer as compared with radiation therapy alone [1,2]. CRT is proposed to be the standard treatment for advanced esophageal cancer [3]. CRT for advanced esophageal cancer has been started in April 2003. The purpose of this study was to examine the treatment outcomes of CRT for advanced esophageal cancer in our institution and analyze their prognostic factors
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