Quantitation in two‐dimensional fluorescence difference gel electrophoresis: Effect of protein fixation

Analyzing a large number of unfixed gels in a 2‐D fluorescence difference gel electrophoresis (2‐DIGE) experiment presents a challenge of avoiding variable protein diffusion within and across the comparison groups. The characteristics of protein detection and quantitation in a 2‐D differential in gel fluorescence experiment were compared for gels with and without protein fixation. The current study tests and concludes that when dealing with a large sample size with variable protein diffusion across the 2‐D gel over a period of 2–4 days, it is a preferred choice to fix the gel without affecting the protein quantitation.