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.
Tannu
Nilesh
N
Department of Physiology and Pharmacology, Wake Forest University School of Medicine, Winston-Salem, NC 27501, USA.
Hemby
Scott E
SE
D001923:Brain Chemistry; D019970:Cocaine-Related Disorders / Q000378:metabolism; D004058:Diffusion; D062787:Drug Overdose / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005404:Fixatives; D005453:Fluorescence; D005456:Fluorescent Dyes; D005782:Gels; D006801:Humans; D009419:Nerve Tissue Proteins / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D015203:Reproducibility of Results
D005404:Fixatives; D005456:Fluorescent Dyes; D005782:Gels; D009419:Nerve Tissue Proteins; D011506:Proteins