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Quantitation in two‐dimensional fluorescence difference gel electrophoresis: Effect of protein fixation
Author(s) -
Tannu Nilesh,
Hemby Scott E.
Publication year - 2006
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200500710
Subject(s) - fluorescence , chemistry , chromatography , difference gel electrophoresis , gel electrophoresis of proteins , gel electrophoresis , two dimensional gel electrophoresis , electrophoresis , color marker , polyacrylamide gel electrophoresis , biochemistry , proteomics , enzyme , physics , gene , quantum mechanics
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.