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A two dimensional electrophoresis database of a human Jurkat T‐cell line
Author(s) -
Thiede Bernd,
Siejak Frank,
Dimmler Christiane,
Jungblut Peter R.,
Rudel Thomas
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(20000701)21:13<2713::aid-elps2713>3.0.co;2-u
Subject(s) - peptide mass fingerprinting , jurkat cells , mass spectrometry , chemistry , matrix assisted laser desorption/ionization , isoelectric point , bottom up proteomics , isoelectric focusing , chromatography , mass spectrum , gel electrophoresis , protein mass spectrometry , peptide , tandem mass spectrometry , proteomics , biology , t cell , desorption , biochemistry , genetics , enzyme , gene , immune system , organic chemistry , adsorption
About 2000 protein spots of human Jurkat T-cells were detected by high resolution two-dimensional gel electrophoresis (2-DE) and were characterized in terms of their isoelectric point and molecular mass. A 2-DE database was constructed and is available at http://www.mpiib-berlin.mpg.de/2D-PAGE/. At present the database contains 67 identified protein spots. These proteins were identified after tryptic digestion by peptide mass fingerprinting with delayed extraction-matrix assisted laser desorption/ionization-mass spectrometry (DE-MALDI-MS). Proteins with a sequence coverage of at least 30% were introduced in the database. This sequence coverage could not always be obtained by using only the matrix alpha-cyano-4-hydroxycinnamic acid (CHCA) for the mass analysis. Therefore, an additional mass spectrum was recorded by using 2,5-dihydroxybenzoic acid (DHB). Usually, additional mass peaks were detected and together with the mass spectrum of CHCA this resulted in the desired sequence coverage.