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“Spot transfer”, elution and comigration with known proteins allows accurate transferral of protein identifications between distinct two‐dimensional electrophoretic systems
Author(s) -
Dean David P.,
Cronan Melissa T.,
Merenda Joseph M.,
Gardner Joseph P.,
Connelly Patricia A.,
Celis Julio E.
Publication year - 1994
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.1150150172
Subject(s) - isoelectric focusing , elution , chromatography , gel electrophoresis , electrophoresis , two dimensional gel electrophoresis , lysis , chemistry , microbiology and biotechnology , biology , biochemistry , proteomics , enzyme , gene
We report a simple method, designated “spot transfer”, where known proteins are excised and eluted from a two‐dimensional (2‐D) gel run on one gel system to transfer identification of the proteins to a different 2‐D gel system by comigration with a comparable sample. In one experiment, 8 of 16 proteins eluted from an isoelectric focusing (IEF) 2‐D gel, of the format described for the Celis human keratinocyte database (Celis, J. E. et al., Electrophoresis 1993, 14 , 1091–1198), were found to comigrate with proteins in a human T lymphoma (JURKAT) whole cell lysate run using the Millipore Investigator 2‐D system. The method should have general utility in allowing the exchange of protein identifications between investigators and could be used to standardize gel loci used in 2‐D gel protein databases.