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Horizontal comparative fluorescence two‐dimensional gel electrophoresis for improved spot coordinate detection
Author(s) -
Hanneken Marina,
König Simone
Publication year - 2014
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.201300507
Subject(s) - difference gel electrophoresis , fluorescence , proteome , chromatography , reproducibility , chemistry , electrophoresis , two dimensional gel electrophoresis , image warping , residual , proteomics , analytical chemistry (journal) , biological system , computer science , biology , artificial intelligence , biochemistry , optics , physics , algorithm , gene
Vertical comparative 2D fluorescence gel electrophoresis (CoFGE) has recently been shown to increase the reproducibility of coordinate assignment for protein spots, in particular in singular experiments, which cannot be investigated using DIGE. The method applies a standardized marker grid formed by a set of purified proteins to the sample proteome in a conglomerate of 1DE, 2DE, and DIGE. Here, improvements are demonstrated by transferring CoFGE to horizontal 2DE. These include the elimination of the protein modification by residual acrylamide monomer unavoidable in vertical CoFGE, reduced buffer volumes, and highly efficient laboratory procedures. Spot patterns are well defined and can be easily analyzed using commercially available warping algorithms. With horizontal CoFGE also a correction for changes in p I was introduced using a third fluorescent dye. Horizontal CoFGE holds high promises in comparative proteomics.