Identification of platelet proteins separated by two-dimensional gel electrophoresis and analyzed by matrix assisted laser desorption/ionization-time of flight-mass spectrometry and detection of tyrosine-phosphorylated proteins.
作者： Katrin Marcus , Dorian Immler , Joachim Sternberger , Helmut E. Meyer
DOI: 10.1002/1522-2683(20000701)21:13<2622::AID-ELPS2622>3.0.CO;2-3
关键词:
摘要: Different search programs were compared to judge their particular efficiency in protein identification. We established a human blood platelet map and identified tyrosine-phosphorylated proteins. The cytosolic fraction of platelets was separated by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) phosphorylated proteins detected Western blotting using anti-phosphotyrosine antibodies. Visualized spots excisted, digested with trypsin analyzed matrix assisted laser desorption/ionization-time flight-mass spectrometry (MALDI-TOF-MS). obtained mass fingerprint data sets have been ProFound, MS-Fit Mascot. For those no significant results MALDI post source decay (PSD) spectra acquired on the same sample. automatic interpretation these fragment ion spectra, SEQUEST Mascot algorithm applied. Another approach for identification is immunoprecipitation an antibody. A method peptides optimized.
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