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An approach for identification of phosphoproteins using the G‐electrode‐loading method in two‐dimensional gel electrophoresis
Author(s) -
Koga Kaori,
Minohata Toshikazu
Publication year - 2011
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201000314
Subject(s) - electrode , chromatography , gel electrophoresis , spots , two dimensional gel electrophoresis , chemistry , materials science , analytical chemistry (journal) , proteomics , microbiology and biotechnology , biology , biochemistry , gene
We assessed whether the G‐electrode‐loading method (GELM) was helpful in the protein analysis. GELM in 2‐DE was compared with the slip‐loading, the in‐gel rehydration and the cup‐loading in 2‐DE. GELM showed the best results for protein separation. A total of 14 spots that showed an increase with GELM were analyzed by MALDI‐TOF MS. In GELM, all of these spots were identified with a high score and a high sequence coverage. A membrane‐associated protein was identified and determined to have phosphorylated site. These tests show that GELM has several advantages for protein analysis compared with the traditional methods.